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Objective:To explore the relationship between cervical microecology and cervical squamous intraepithelial lesions (SIL).Methods:All subjects were recruited from the health care center or gynecology of the Affiliated Wuxi No.2 People′s Hospital of Nanjing Medical University from March to May of 2019, including 12 subjects normal cervix with 37-47 years old, 21 low-grade squamous intraepithelial lesion (LSIL) subjects with 39-48 years old, 5 high-grade squamous intraepithelial lesion (HSIL) subjects with 38-45 years old and 3 cervical squamous cell carcinoma subjects with 42-43 years old. All subjects were required to fill in a questionnaire, and performed cervical examination. Meanwhile, the microecology of cervical secretions was analyzed by the next generation sequencing (NGS) and the NGS results were analyzed by bioinformatics. Subjects were divided into human papilloma virus (HPV)-negative groups, low-risk HPV (lrHPV), 16/18 high-risk HPV (hrHPV) and other hrHPV infection groups based on HPV test results of NGS. The Venn diagram of data, microecology diversity, the relative abundance and co-occurrence of species, and the receiver operating characteristic (ROC) curve were analyzed.Results:A total of 909 species at the species level were obtained from the cervical secretions of all the subjects, and there was overlap among the groups. There was no significant difference in total HPV infection rate, 16/18 hrHPV infection rate and other hrHPV infection rates among subjects with different cervical lesions (all of P>0.05). Grouped by HPV infection, the 16/18 hrHPV-infected and other hrHPV-infected subjects had increased cervical microecology diversity ( U=39.00 and 43.00, all of P<0.05), and the relative abundance of Lactobacillus crispatus (L.crispatus) had no differences among the groups ( H=4.37, P=0.213 6). Grouped by cervical conditions, the cervical microecology diversity of the subjects with cervical lesions increased ( H=14.60, P=0.002 2), while the L.crispatus relative abundance decreased ( H=13.98, P=0.000 8). Among all the detected species, Mycoplasma, Chlamydia and Streptococcus B had a co-occurrence, while Lactobacillus iners, Gardnerella vaginalis, Atopobium vaginae, and Prevotella bivia had a co-occurrence. As the SIL diagnostic index, the area under the ROC curve (AUC) of the relative L.crispatus relative abundance was 0.874 [95% confidence interval ( CI):0.732-0.957]. L.crispatus combined with Lactobacillus jensenii (L.jensenii) and Mycoplasma had an AUC of 0.943 [95 %CI: 0.822-0.991] in the SIL diagnosis. Conclusions:The decreased L.crispatus relative abundance and the increased cervical microecology diversity may be related to HPV infection and cervical lesions; simplified NGS data may be helpful to the SIL diagnosis.
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Objective@#To establish a Galleria mellonella model of liver abscess-related Klebsiella pneumoniae (K.pneumoniae) infection and to evaluate its feasibility for virulence detection.@*Methods@#Twelve liver abscess-related K. pneumoniae strains were collected in Wuxi No.2 People′s Hospital from January 2016 to December 2017. Twenty K. pneumoniae strains isolated from sputum and urine samples were used as classic strains. All isolates were analyzed by String test. Common capsular serotypes (K1, K2, K5, K16, K20, K54 and K57) of highly virulent K. pneumoniae strains were detected by PCR. Virulence test was performed to measure 80% lethal doses (LD80) of different serotypes in the same time period and the lethal time for 80% of larvae (LT80) at the same concentration.@*Results@#The 12 strains of liver abscess-related K. pneumoniae belonged to five serotypes, which were K1 (41.7%, 5/12), K2 (8.3%, 1/12), K5 (8.3%, 1/12), K20 (8.3%, 1/12) and K57 (33.4%, 4/12). High-virulence serotypes were not detected in the classic group. The positive rates of String test in the liver abscess group and the classic group were 75% and 10% (2/20), respectively. Results of the virulence test showed that when the concentration ranged from 1×105 CFU/ml to 1×108 CFU/ml, the lethal effects of different strains on Galleria mellonella larvae were in a concentration dependent manner. Twelve hours after infection, the numbers of dead larvae in K1 and K57 serotype groups were significantly higher than those in K2, K5, K20 and classic groups. The LD80 values of the liver abscess group at 96 hours after infection were as follows: 1×106 CFU/ml (K1, K57) and 1×107 CFU/ml (K2, K5, K20).@*Conclusion@#The liver abscess-related K. pneumoniae isolates are all highly virulent strains. The virulence of K. pneumoniae can be detected at 12 hours after infection of Galleria mellonella.
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Objective To investigate the distribution of hypervirulent capsular serotypes and virulence genes of Klebsiella pneumoniae associated with pyogenic liver abscess(PLA),and analyze the homology of strains.Methods Twelve strains of liver abscess-related Klebsiella pneumoniae were isolated in Wuxi Second People's Hospital during January 2016 and August 2017.Among them,five were also detected in blood samples.All the strains were performed the viscous thread test,and the hypervirulent capsular serotypes and main virulence genes were detected by PCR.The homology of the strains was analyzed by the multilocus sequence types(MLST) and pulsed-field gel electrophoresis(PFGE).Results The positive rate of the viscous thread test for 12 strains of liver abscess-related Klebsiella pneumoniae was 75％.Three kinds of hypervirulent capsular serotypes,including K1 (6/12,50％),K54(1/12,8.3％) and K57(5/12,41.7％),were detected.The positive rates of virulence genes,such as wcaG,rmpA,ureA,fimH,mrkD,uge,Aer and iroNB,were all 100％,while that of iucB was 83.3％.The cf29a gene was not found,and the magA,allS and kfuBC genes were only detected in K1 serotype strains.MLST found that ST23(4/12,33.3％) and ST25(3/12,25％) were main sequence types,and then were ST412(2/12,16.7％),ST1660(1/12,8.3％),ST1049(1/t2,8.3％) and ST11 (1/12,8.3％).PFGE showed that 12 strains of Klebsiella pneumoniae were divided into 8 types,and that only 3 strains of K1 serotype belong to the same clonotype.Conclusion All the isolated liver abscess-related Klebsiella pneumoniae strains are highly virulent.ST23 and ST25 are main sequence types,and ST1049 is first reported.PFGE results show genetic diversity,and Kl-type Klebsiella pneumoniae has a certain epidemic tendency.
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Objective To evaluate the early diagnostic value of the detection of plasma 1,3-β-D glucan(G test) for deep mycobacterium infection and assess the difference of the G test and fungal culture.Methods We collected test plasma from 420 subjects from January to November 2014,including 226 cases of deep mycobacterium infection,88 cases of non-fungal infection and 106 cases of healthy controls.G-test was used to measure the concentration of 1,3-β-D glucan in the plasma.The cut-off points for 1,3-β-D glucan were determined by using receiver operator characteristics curves(ROC).Results The concentration of 1,3-β-D in plasma of paients with deep mycobacterium infection was significantly higher than that of patients with non-fungalinfection and healthy controls.There was no difference between the control groups.With the analysis of ROC curve,the best cut-off value was 25.33 pg/mL.The sensitivity,specificity,positive predictive value and negative predictive value of G test were 84.9%,70.83%,72.0%,84.1%(P<0.05).Conclusion G Test is a practical and effective method in early diagnosis of deep fungal infection.
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Objectives To investigate the mechanism of integron mediated resistance and multidrug-resistance in P.aeruginosa.Methods The variable region of integron was amplified by integron PCR.Restriction fragment length polymorphism(RFLP)and DNA sequencing were used to investigate the resistance genes in the variable region of integron.Results Of the 98 strains 35(35.7%)were the variable region positive,and size of the amplicons ranged from 1.0 kb to 4.0 kb.A total of 6 different cassette arrays were detected,including genes coding resistance to aminoglycosides,?-lactam compounds and sulfanilamides.Of the 5 cassette arrays 3 were novel,including aadA6-orfD,aadB-blaP1 and aadB-aac6-Ⅱ-blaCARB-8,and the Genbank numbers were DQ 091179,DQ 141316 and DQ 288251 respectively.Conclusions Integron mediates the resistance and multidrug resistance in P.aeruginosa.The majority of the genes located in integrons are those coding resistance to aminoglycosides.Three strains of class I integron with novel cassette arrays are reported.