1.Molecular epidemiological analysis of Streptococcus pyogenes strains isolated from the adult and pediatric patients in Shanghai,China
Haoqin JIANG ; Mingliang CHEN ; Tianming LI ; Hong LIU ; Mei ZENG ; Min LI
Chinese Journal of Infection and Chemotherapy 2015;(3):222-229
Objective To investigate and compare the molecular epidemiological characteristics of Streptococcus pyogenes isolates from Shanghai adult and pediatric patients in terms of antimicrobial susceptibility ,clone type ,emm type ,biofilm formation and virulence for better infection control and treatment .Methods Thirty‐nine nonduplicate clinical isolates of S . pyogenes from adult and pediatric patients were analyzed by determining the susceptibility to antimicrobial agents by Kirby‐Bauer method;clonal typing by multilocus sequence typing ( MLST ); genotyping by emm gene sequence analysis ,which encoding M protein;genomic characteristics of different emm type strains by pulsed field gel electrophoresis (PFGE );and biofilm formation by semi‐quantitative biofilm formation test . Twenty main virulence genes of S .pyogenes ,including 12 superantigen genes and 8 other key genes were detected by PCR and gel electrophoresis . Results A total of 39 nonduplicate S .pyogenes isolates were analyzed .The most common genotype was emm 12‐ST36 (64 .1% ) and emm 1‐ST28 (17 .9% ) .Isolates from adult and pediatric patients had the same dominant genotype , emm 12‐ST36 . The isolates from children showed significantly higher resistance rate to erythromycin and clindamycin than those from adult patients (P<0 .000 1) .Particular emm type and clone type were frequently identified in the same PFGE cluster .Statistical analysis showed that biofilm formation was significantly associated with emm type 1 (P=0 .005) and erythromycin/clindamycin resistance (P=0 .000 3) .The strains from children showed higher biofilm formation than those from adult patients (P<0 .000 1) .We found that virulence genes speA ,speJ and spd3 were significantly associated with emm type 1 (P<0 .000 1 ,P=0 .005 5 ,P<0 .000 1) ,while speI and sic were significantly associated with emm type 12 (both P<0 .000 1) .We also found that the prevalence of speC ,speH ,ssa , smeZ ,and sdaD genes was significantly different between emm type 12 and emm type 1 (P= 0 .023 8 , P< 0 .000 1 , P<0.0001,P= 0.0003,and P= 0.0068,respectively).TheprevalenceofvirulencegenesspeH,smeZandsdaDwas significantly different between the emm type 12 strains from children and those from adults (all P< 0 .000 1) .Conclusions There is a strong agreement between emm type ,clone type ,virulence genes and the clusters defined by PFGE profiling of S . pyogenes .S .pyogenes isolates from adult and pediatric patients are different in terms of antibiotic resistance and biofilm formation .Certain emm type is significantly associated with antibiotic resistance and virulence ,which is useful for infection control .Dominant virulence genes may be the potential target for developing new vaccine to reduce S .pyogenes infection in the future .
2.Molecular characteristics of Streptococcus pneumoniae in Shanghai, China
Haoqin JIANG ; Mingliang CHEN ; Tianming LI ; Yueru TIAN ; Yuanjun ZHU ; Hong LIU ; Min LI
Chinese Journal of Laboratory Medicine 2015;38(4):256-261
Objective Toinvestigate the molecular characteristics including antibiotic resistance,strain type,serotype,virulence,biofilm formation of Streptococcus pneumoniae isolated from Shanghai adult patients.Methods A total of 37 non-repetitive S.pneumoniae isolates causing community acquired and hospital acquired infections of adults were collected from Shanghai Huashan Hospital from January 2011 to December 2013.The inhibitory zone diameter or minimum inhibitory concentrations (MICs) of 9 antimicrobial agents (penicillin,vancomycin,erythromycin,clindamycin,levofloxacin,cefprozi,ceftriaxone,cefotaxime and linezolid) were determined by Kirby-bauer (K-B) method or Etest method;Serotypes were tested by polymerase chain reaction (PCR) and S.pneumoniae antisera agglutination;Genomic characteristics of different serotype strains were determined by pulsed field gel electrophoresis (PFGE)method;Multilocus sequence types (MLST) was used for strain type;Semi quantitative biofilm formation test was used for the biological membrane formation.Ten main pneumococcal virulence genes (cbpA,pspA,cps2A,lytA,nana,pavA,piaA,ply,psaA and spxB) were detected by PCR and gel electrophoresis.Statistical analysis was performed using Stata software and association statistics were tested using Fisher's exact test.Results The most frequent serotypes were 19F (13.5%),23 F (13.5%),14 (10.8%),19A (10.8%).The penicillin resistance rate was 64.9%.Serotypes 19 F,19A and 23 F were significantly associated with penicillin resistance (x2 =5.89,P =0.015) and the isolates belonged to these serotypes were all multi-drug resistant (MDR).ST81 and ST271 showed high resistance rates to several antibiotics including penicillin (x2 =4.57,P =0.033).Biofilm formation was significantly associated with serotypes 19A (x2 =5.55,P =0.018) and strain type ST320 (x2 =4.33,P =0.037),but not associated with penicillin resistance (x2 =0.16,P =0.686).Virulence gene lytA,pavA,ply,psaA,spxB were found in all isolates.Conclusions Penicillin resistance rate of S.pneumoniae in adult is rising.Specific serotype,epidemic clone and antibiotic resistance are closely related,and can provide the basis for the infection control.The virulence factors such as PspA will be the new targets for vaccine development to reduce S.pneumoniae infection in the future.
3.The study of SALT classification method for health service support in disaster assistance
Yucui ZHAI ; Dongmei CHEN ; Haoqin REN ; Ling DAI
Chinese Journal of Practical Nursing 2018;34(17):1296-1300
Objective To study the value of SALT method in disaster assistance. Methods A total of 84 internship nurses from 2015 to 2017 were randomly divided into group SLAT and group START and accepted different classification methods training, then checked under the scene of simulated mass casualty events (car accident and earthquake) and the results of the examination were compared. Results The triage time of the SALT group was (8.00±1.36) min, (23.14±1.95) min,less than (10.05±1.34) min, (25.07±2.22) min in the START group in the car accident scene and the earthquake scene and the differences were statistically significant (t=-6.94,-4.23, P<0.05). The accuracy of different classifications under the same scene were compared and there was no statistical significance in the difference(P>0.05). The accuracy of the triage in the car accident scene were better than earthquake scenes of the both SALT and START classifications and differences were statistically significant ( χ2=10.49, 10.54,P<0.05). The consistency of the SALT classification and the START classification was better (Kappa=0.526, P<0.01). Conclusions The SALT method has the advantages of simple, easy to master, accurate and reliable. It is consistent with the traditional START method, but the time of classification is shorter and recommended to be used in the teaching of disaster medicine and the rescue of real disasters.
4.Test of Infant Motor Performance in the developmental assessment of preterm infants
Sa YUAN ; Haoqin ZHOU ; Huiping ZHANG ; Haiyan YING ; Ru JIAN ; Yanni CHEN
Chinese Journal of Applied Clinical Pediatrics 2023;38(2):120-124
Objective:To assess the characteristics and correlation of motor development in preterm infants of different gestational weeks by using the Test of Infant Motor Performance (TIMP) method, and to develop better individualized early interventions based on TIMP test results.Methods:A prospective study involving 43 full-term healthy infants and 77 preterm followed up in 3201 Hospital from June 2019 to July 2021 was conducted.Preterm infants were divided into the early preterm group (39 cases) and late preterm group (38 cases) according to their gestational age at birth.TIMP assessment was performed at the gestational age of 40 weeks and the corrected age of 16 weeks after birth.Similarly, the full-term healthy infants were assessed by TIMP at the postnatal age of 16 weeks.The differences between groups were investigated using ANOVA or Mann- Whitney rank sum test.Correlations were analyzed by the Pearson correlation method. Results:There were no significant difference in TIMP scores between early and late preterm infants at the gestational age of 40 weeks [(65.74±6.52) scores vs.(66.96±8.51) scores] and the corrected age of 16 weeks [(101±10) scores vs.(104±8) scores] (all P>0.05). TIMP scores in the full-term healthy group at the corrected age of 16 weeks [(108±10) scores] differed significantly from those of early and late preterm infants ( P<0.05). Compared with full-term infants, early and late preterm infants had lower TIMP scores in observation, supine position, and supine turning (all P<0.05), but a higher TIMP score in standing position ( P<0.05). For both early and late preterm infants, TIMP scores at the gestational age of 40 weeks were significantly positively correlated with those at the corrected age of 16 weeks ( r=0.565, 0.302, all P<0.01). Conclusions:There were significant differences in motor development between preterm infants of different gestational ages and term infants, which had guiding significance for early intervention.English version TIMP could play a positive role in promoting individualized follow-up and early intervention of preterm infants in China.
5.A multi-center study on evaluation of leukocyte differential performance by an artificial intelligence-based Digital Cell Morphology Analyzer
Haoqin JIANG ; Wei CHEN ; Jun HE ; Hong JIANG ; Dandan LIU ; Min LIU ; Mianyang LI ; Zhigang MAO ; Yuling PAN ; Chenxue QU ; Linlin QU ; Dehua SUN ; Ziyong SUN ; Jianbiao WANG ; Wenjing WU ; Xuefeng WANG ; Wei XU ; Ying XING ; Chi ZHANG ; Lei ZHENG ; Shihong ZHANG ; Ming GUAN
Chinese Journal of Laboratory Medicine 2023;46(3):265-273
Objective:To evaluate the performance of an artificial intelligent (AI)-based automated digital cell morphology analyzer (hereinafter referred as AI morphology analyzer) in detecting peripheral white blood cells (WBCs).Methods:A multi-center study. 1. A total of 3010 venous blood samples were collected from 11 tertiary hospitals nationwide, and 14 types of WBCs were analyzed with the AI morphology analyzers. The pre-classification results were compared with the post-classification results reviewed by senior morphological experts in evaluate the accuracy, sensitivity, specificity, and agreement of the AI morphology analyzers on the WBC pre-classification. 2. 400 blood samples (no less than 50% of the samples with abnormal WBCs after pre-classification and manual review) were selected from 3 010 samples, and the morphologists conducted manual microscopic examinations to differentiate different types of WBCs. The correlation between the post-classification and the manual microscopic examination results was analyzed. 3. Blood samples of patients diagnosed with lymphoma, acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndrome, or myeloproliferative neoplasms were selected from the 3 010 blood samples. The performance of the AI morphology analyzers in these five hematological malignancies was evaluated by comparing the pre-classification and post-classification results. Cohen′s kappa test was used to analyze the consistency of WBC pre-classification and expert audit results, and Passing-Bablock regression analysis was used for comparison test, and accuracy, sensitivity, specificity, and agreement were calculated according to the formula.Results:1. AI morphology analyzers can pre-classify 14 types of WBCs and nucleated red blood cells. Compared with the post-classification results reviewed by senior morphological experts, the pre-classification accuracy of total WBCs reached 97.97%, of which the pre-classification accuracies of normal WBCs and abnormal WBCs were more than 96% and 87%, respectively. 2. The post-classification results reviewed by senior morphological experts correlated well with the manual differential results for all types of WBCs and nucleated red blood cells (neutrophils, lymphocytes, monocytes, eosinophils, basophils, immature granulocytes, blast cells, nucleated erythrocytes and malignant cells r>0.90 respectively, reactive lymphocytes r=0.85). With reference, the positive smear of abnormal cell types defined by The International Consensus Group for Hematology, the AI morphology analyzer has the similar screening ability for abnormal WBC samples as the manual microscopic examination. 3. For the blood samples with malignant hematologic diseases, the AI morphology analyzers showed accuracies higher than 84% on blast cells pre-classification, and the sensitivities were higher than 94%. In acute myeloid leukemia, the sensitivity of abnormal promyelocytes pre-classification exceeded 95%. Conclusion:The AI morphology analyzer showed high pre-classification accuracies and sensitivities on all types of leukocytes in peripheral blood when comparing with the post-classification results reviewed by experts. The post-classification results also showed a good correlation with the manual differential results. The AI morphology analyzer provides an efficient adjunctive white blood cell detection method for screening malignant hematological diseases.
6.The lncSIL molecule exerts a negative regulatory effect on the alveolar epithelial-mesenchymal transition induced by TGF-β1 through modulation of the EZH2/P21/CDK6 signaling pathway
Wanfang Zhang ; Lin Wang ; Pengtao Pan ; Wenxin Li ; Ruili Kang ; Ziren Zhu ; Haoqin Chen ; Xinyu Fang ; Xingcan Zhang ; Yuxin Zhang ; Yiwen Jiang ; Xinyan Li ; Benqi Yuan
Acta Universitatis Medicinalis Anhui 2024;59(4):600-604
Objective :
To investigate the role of lncSIL in transforming growth factor-β1(TGF-β1)-induced alveo- lar epithelial interstitial transformation (EMT) and its related signaling pathways .
Methods :
Western blot was used to detect the effect of lncSIL silencing on the expression of E-cadherin ( E-cad) , alpha-smooth muscle actin ( α- SMA) and Collagen I (Col I) in the process of EMT induced by TGF-β1 . LncSIL interacting proteins were ana- lyzed by RNA pulldown . Western blot was used to detect the effect of overexpression or silencing of lncSIL on the expression of its target gene enhancer of zeste homolog 2 (EZH2) and its downstream factors P21 and cyclin-de- pendent kinase 6 (CDK6) . Flow cytometry was used to analyze the effect of lncSIL on cell cycle progression .
Results:
After lncSIL silencing , the expression of α-SMA and Col I increased , the expression of E-cad decreased . RNA pulldown assay showed that EZH2 was the target protein that interacted with lncSIL , and the expression of EZH2 increased after silencing lncSIL , the expression of EZH2 downstream gene P21 decreased , CDK6 increased . Flow cytometry showed that the number of cells in S phase significantly increased . When lncSIL was overexpressed , the expression of EZH2 and CDK6 was down-regulated , the expression of P21 was up-regulated , and the number of S phase cells significantly decreased .
Conclusion
LncSIL inhibits TGF-β1-induced alveolar epithelial cell mesen- chymal transition by negatively regulating EZH2/P21 /CDK6 signaling pathway to inhibit cell cycle progression .