3.Direct identification of pathogens from positive blood cultures by matrix-assisted laser desorption ionization-time of flight mass spectrometry
Xiaofang XIE ; Huiqin ZHOU ; Yi ZHENG ; Tingting LIU ; Rongqin XU ; Hong DU
Chinese Journal of Clinical Infectious Diseases 2016;9(2):152-155
Objective To develop a rapid and accurate technique for pathogens identification from positive blood cultures by matrix-assisted laser desorption ionization-time of flight mass spectrometry ( MALDI-TOF MS) .Methods A total of 266 culture-positive blood samples from clinical laboratory in the Second Affiliated Hospital of Soochow University during January to July 2015 were collected.The blood was transferred into separation-gel tubes, and the bacteria were enriched and purified by differential centrifugation.MALDI-TOF MS was applied to identify the bacteria and the results were compared with conventional bacterial culture. Results Among 266 culture-positive blood samples, 260 were monomicrobial cultures and 6 were polymicrobial cultures.Of 260 monomicrobial cultures, 98.8% (257/260) and 96.2% ( 250/260 ) of organisms were identified at the genus level and the species level, respectively.Of 140 Gram-negative bacterial isolates, 99.3% (139/140) and 97.9% (137/140) were identified at the genus level and the species level, respectively.Of 120 Gram-positive bacteria isolates, 98.3%(118/120) and 94.2% (113/120) were identified at the genus level and the species level, respectively.None of the 6 polymicrobial cultures were identified.Conclusion MALDI-TOF MS can directly identify the bacteria from positive blood cultures, which provides a rapid and accurate method for the diagnosis of bloodstream infection.
4.Study on Quality Control of New Medicinal Resources of Pineapple Leave
Jingfei JIANG ; Wei WANG ; Zhenguo LI ; Fan LEI ; Weidong XIE ; Yi DING ; Dongming XING ; Lijun DU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):972-977
This article was aimed to study the quality control of pineapple leaf in order to further develop the appli-cations of pineapple leave as medicinal resources. The tissue structure of pineapple leaf was observed by tissue biop-sy assay. Thin-layer chromatography (TLC) was employed on the qualitative identification of ananasate in the leave. The p-coumaric acid and ananasate were detected with high-performance liquid chromatography (HPLC) assay in the content determination. The results showed that pineapple leaf can be divided into the upper and lower leaf epidermis with stoma under microscope observation. Ananasate of the leave can be indentified clearly by TLC. HPLC method can accurately detect the content of p-coumaric acid and ananasate of pineapple leave. There are different contents of p-coumaric acid and ananasate of pineapple leave in different origins. In Y unnan province, the contents of p-coumaric acid and ananasate of pineapple leave are the highest; while the contents of p-coumaric acid and ananasate in Guangxi province are the lowest. It was concluded that ananasate can be used in the quality identification. The p-coumaric acid and ananasate can be used in the quantitative determination in the better control of the quality of pineapple leaf.
5.Cytogenetic abnormalities detected by fluorescence in situ hybridization panel in chronic lymphocytic leukemia
Yi YING ; Xiaoyan CHEN ; Qinghua DU ; Hanping WANG ; Jianjin XIE ; Xiaowei CHEN
Journal of Leukemia & Lymphoma 2014;23(7):416-419
Objective To investigate the significance of fluorescence in situ hybridization (FISH) panel in detecting cytogenetic abnormalities in patients with chronic lymphocytic leukemia (CLL).Methods A panel of FISH probes [D13S25 (13q14.3),RB1 (13q14),ATM(1 1q22.3),CSP12(12p1 1.1-12q1 1.1) and p53 (17p13.1)] were performed in 21 cases with CLL.The cytogenetic features in correlation with clinical manifestation,other laboratory tests and prognosis were analyzed.Results Cytogenetic abnormalities were found in 13 of 21 patients with CLL (61.90 %).The most frequent abnormality was del(13q14) (42.86 %),followed by trisomy 12 (14.29 %),del(11q22) (9.52 %) and del(17p13) (9.52 %).There was no significant relationship among cytogenetic abnormalities and sex,binet stages,expression of CD38,level of lactate dehydrogenase.Conclusion FISH with probe panel is a rapid,sensitive and accurate technique for detection of cytogenetic abnormalities in patients with CLL.
6.Study on effect of Chuankezhi in assisted treatment of capillary bronchitis
Jiyu DU ; Yaqiong LIU ; Jia CHEN ; Jian XIE ; Jing YU ; Yi ZHANG
Chongqing Medicine 2017;46(22):3085-3087
Objective To observe the clinical curative effect of Chuankezhi Injection atomization inhalation in treating capillary bronchitis.Methods 93 ninety-three children cases of capillary bronchitis were in our hospital from January to December 2015were selected and randomly divided into the treatment group(50 cases) and control group(43 cases) according to the random number table method.The two groups were given the same routine treatment.The treatment group was simuhaneously given 1 mL of Chuankezhi Injection adding into 4 mL of 0.9% Sodium Chloride Injection,the mixture was inhaled by oxygen atomization at a speed of 4-5 L/s for 10-12 min,with 7 d as one treatment course.The clinical effect,clinical symptom disappearance situation and hospitalization time in the two groups were observed.Results The total effective rate in the treatment group was 98.0% (49/50),which was higher than 79.1% (34/43) in the control group;the disappearance time of cough,dyspnea and pulmonary wheezes and crackles and rales,and hospitalizatiorn time in the treatment group were (6.15 ± 1.50)d,(4.5 ± 1.90)d,(4.60 ± 1.70)d,(5.52 ±1.31)d and (6.55±2.30) d respectively,which were significantly better than (7.19 ± 1.85) d,(5.7 ± 2.10) d,(5.81 ± 1.82) d,(6.50 ± 1.83)d and (7.48 ± 2.51) d in the control group,the differences of various indexes between the two groups were statistically significant(P<0.05).Conclusion Chuankezhi Injection in the assisted treatment of capillary bronchitis has significantly effect and better clinical application value.
7.Application of the Peak Area Ratio of STR Loci to Amelogenin Locus in the Estimation of DNA Degradation.
Ya-ling XIE ; Lu LI ; Cheng-chen SHAO ; Yi-hui WU ; Tie-shuai DU ; Huai-gu ZHOU ; Hui LI ; Jian-hui XIE ; Yi-wen SHEN
Journal of Forensic Medicine 2016;32(2):105-108
OBJECTIVE:
To explore the change rules of peak area ratio of STR loci to Amelogenin (AMEL) locus (STR/AMEL), a sex-determining gene in DNA degradation, and to evaluate the application of STR/AMEL value in the estimation of DNA degradation degree.
METHODS:
DNA was extracted from iliopsoas, and the variations of STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) were analyzed after the artificial degradation was made by DNase I, and the changes of these three ratios of the iliopsoas naturally degraded in an outdoor environment were also analyzed. The regression curves were analyzed using the periods of DNA degradation and outside the body as the independent variable (x) and the STR/AMEL value as the dependent variable (y) and three curve equations under two conditions were established.
RESULTS:
Both under the conditions of artificial and natural degradation, STR/AMEL value had a negative relationship with the degradation time. The relationship between STR/AMEL and degradation time can be well simulated by the cubic function. R2 was over 0.99 under controlled degradation condition and over 0.86 under natural degradation condition.
CONCLUSION
The STR/AMEL value (Penta E/AMEL, Penta D/AMEL, FGA/AMEL) is negatively related with the DNA degradation degree, which follows mathematical regression models strictly, and it might be applied to evaluate the DNA degradation degree.
Amelogenin/genetics*
;
DNA Damage/genetics*
;
DNA Primers
;
Humans
;
Microsatellite Repeats
;
Regression Analysis
;
Time Factors
8.The preliminary study of CD4+ Vα9-J27/Vβ29-D1-J2 tetramers in detecting Mycobacterium tuberculosis infections
Dan XIE ; Kouxing ZHANG ; Xuanjing DU ; Yimin FANG ; Yan LI ; Yi CHEN ; Jianbo ZHANG ; Ming GAO ; Xiaomin LAI
Chinese Journal of Microbiology and Immunology 2012;32(1):20-24
ObjectiveTo investigate the specificity of CD4+ Vα9-J27/Vβ29-D1-J2 tetramer in detecting Mycobacterium tuberculosis(MTB) infections.MethodsThe above TCR tetramer by using biotinylated monomers expressed and purified from constructed stable Drosophila Schneider 2 cell( S2 cell) lines was prepared.The PE-labled TCR tetramer was used to costain with S2 cell lines expressing MTB prptide/HLA-DR complexes on the cell membrane,and also was used to detect tetramer-bound CD14+ monocytes and macrophages in the peripheral blood mononuclear cells (PBMC) of pulmonary tuberculosis (PTB) patients and three control groups by flow cytometric analysis.And the FITC-labled tetramer was used to examine tetramer-bound CD14+ monocytes and macrophages,and MTB antigen-specific and tetramer-bound cells by in situ staining.ResultsThe TCR tetramer was well binding with S2 cell lines expressing C14/HLA-DR *1504 on the cell membrane.By flow cytometric analysis,the percentage of tetramer-bound CD14+ monocytes and macrophages in PTB patients group was higher than the other three control groups( P<0.001 ).By in situ staining,tetramer-bound CD14+ monocytes and macrophages,and MTB antigen-specific and tetramer-bound cells were positive in PTB tissue and negative in control pneumonia tissue.ConclusionThe spcificity of TCR tetramer in monitoring MTB infections by flow cytometric analysis and in situ staining could be seen,which laid a laboratory foundation in the diagnosis and immune mechanism research of TB by using TCR tetramers.
9.The value of quantitative artificial intelligence measurement in evaluation of CT dynamic changes for COVID-19
Dan DU ; Yuanliang XIE ; Hui LI ; Shengchao ZHAO ; Yi DING ; Pei YANG ; Bin LIU ; Jianqing SUN ; Xiang WANG
Chinese Journal of Radiology 2021;55(3):250-256
Objective:To investigate the value of artificial intelligence (AI)-assisted quantitative measurement in evaluation of the dynamic changes of CT for COVID-19 pneumonia.Methods:The clinical and chest CT dynamic imaging data of 99 patients with confirmed COVID-19 pneumonia who were hospitalized in Wuhan Central Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 15, 2020 to March 10, 2020 were retrospectively analyzed. According to the definitive diagnosis, the 99 patients were classified into common ( n=36), severe ( n=33) and critical ( n=30) type, the CT imaging findings of each type were analyzed, including CT basic signs, total volume of pneumonia lesions and percentage of pneumonia lesions of the total lung volume (volume ratio). AI software was used to quantitatively evaluate the dynamic changes of chest CT images. The quantitative indicators included CT peak time of lesions, total volume of lesions peak, volume ratio of lesions peak, maximum growth rate of total volume and maximum growth rate of volume ratio. Kruskal-Wallis rank sum test was used to compare the difference of quantitative indexes between the 3 types, and χ 2 test or Fisher exact probability test was used to compare the difference of qualitative indexes between the 3 types. Sequence measurement and scatter plots were used to show the evolution trend of the volume ratio of the three types of COVID-19 pneumonia lesions. The ROC curve was used to analyze the value of the volume ratio of pneumonia lesions and its maximum growth rate in predicting the conversion of common pneumonia to severe or critical pneumonia. Results:There were statistically significant differences in age and gender distribution among patients with common, severe and critical COVID-19 ( P<0.05), the age of severe and critical types were significantly higher than that of common type ( P<0.01). Compared with common [2.5 (1.0, 5.0) d] and critical type[2.5 (1.0, 4.0) d], the time from onset to the first chest CT scan of severe type was prolonged [5.0 (2.5, 8.0) d, P<0.01]. There were statistically significant differences in involvement of multiple lung lobes (20 cases, 29 cases, 25 cases, χ2=10.403, P=0.006) in patients with common, severe and critical COVID-19 at the first scan, the incidence of the involvement of multiple lung lobes in severe and critical types was significantly higher than that of common type ( P=0.002). The volume ratios of patients with common, severe and critical COVID-19 at the first scan were statistically significant [1.0% (0.2%, 4.7%), 9.30% (1.63%, 26.83%), 2.10% (0.64%, 8.61%), Z=14.236, P=0.001], and the volume ratio of severe type was significantly higher than that of common type ( P<0.001), there was no statistically significant difference between common type and critical type ( P=0.062). Follow-up CT showed that the pneumonia lesions showed a dynamic transformation of progress and recovery, and it was seen that the coexistence of multiphase lesions. The trend line in the scatter plot of the three types of COVID-19 pneumonia lesions showed that the lesions in the advanced stage developed from less to more. The lesion peak volume ratios of the common, severe and critical types were 9.75% (4.83%, 13.18%), 29.80% (23.99%, 42.36%) and 61.81% (43.73%, 72.82%), respectively, the difference was statistically significant ( Z=74.147, P<0.001). The maximum growth rates of lesion volume ratio were 1.27% (0.50%, 1.81%)/d, 4.39% (3.16%, 5.54%)/d and 6.02% (4.77%, 9.96%)/d, respectively, the difference was statistically significant ( Z=52.453, P<0.001). The peak times of lesions were 12.0 (9.0, 15.0) d, 13.0 (10.0, 16.0) d and 16.5 (12.0, 25.0)d, respectively, the difference was statistically significant ( Z=9.524, P=0.009). Taking the volume ratio of pneumonia lesion 22.60% and the maximum growth rate of the volume ratio 1.875%/d as the boundary value, the sensitivity of diagnosing common type to severe or critical type was 92.10% and 96.83%, and the specificity was 100% and 80.56%, respectively. The area under the curve was 0.987 and 0.925, respectively. Conclusions:The lesions of COVID-19 pneumonia show a similar parabolic change on CT imaging. The use of AI technology to dynamitcally and accurately measure the CT pneumonia lesion volume ratio is helpful to evaluate the severity of the disease and predict the development trend of the disease. Patients with a rapid growth of volume ratio are more likely to become severe or critical type.
10.Analysis of the hemolysin and virulence-related genes of incomplete hemolysis Staphylococcus aureus
Yi ZHENG ; Xiaofang XIE ; Hong DU ; Haifang ZHANG ; Xueming ZHU ; Min WANG ; Huan YANG ; Aiqing LI ; Ye JIANG ; Huiqin ZHOU
Chinese Journal of Clinical Infectious Diseases 2016;9(3):236-242
Objective To analyze hemolysin and virulence -related genes in incomplete hemolytic Staphylococcus aureus.Methods Fifty strains of incomplete hemolytic Staphylococcus aureus were isolated from patients admitted in the Second Affiliated Hospital of Soochow University during 2013 and 2014, and the isolates with complete hemolytic phenotype were also collected at the same period as the control strains . All the strains were inoculated and subcultured on four kinds of sheep blood agar plates supplied by different manufacturers to compare their hemolytic phenotype .The relative mRNA expressions of hemolysin genes (hla, hlb, hlc, hld) in standard strain, complete and incomplete hemolytic phenotype strains were detected by real-time quantitative polymerase chain reaction (RT-qPCR), and valued by 2 -△△Ct method.t test was used to compare mRNA expressions of hemolysin genes .Western blot was performed to analyze the expression of α-hemolysin.Antibiotic susceptibility test of incomplete hemolytic strains was performed using broth microdilution method.Resistant gene mecA and virulence genes pvl, tst were detected by PCR.Results The steady and hereditary incomplete hemolysis was observed in 50 strains of incomplete hemolytic Staphylococcus aureus on the sheep blood agar plates from different suppliers .Taking mRNA expression of hla, hlb, hlc, hld in standard strain as 1, the relative mRNA expressions of hemolysin genes in incomplete hemolytic strains were 0.02, 7.51, 0.06 and 0.12 respectively, there were statistical differences between standard strain and incomplete hemolytic strains (t =8.46, -56.40, 8.12 and 7.61, all P <0.05).And the expression of α-hemolysin was decreased in incomplete hemolytic strains .All the strains were identified as methicillin resistant Staphylococcus aureus (MRSA).Three strains exhibited different minimum inhibitory concentrations of teicoplanin and linezolid after subcultured , but the differences had no impact on the final results of antibiotic susceptibility test .mecA, pvl and tst genes were positive in incomplete hemolytic strains . Conclusion Staphylococcus aureus with incomplete hemolytic phenotype is methicillin resistant with higher expression of β-hemolysin and lower expressions of α-hemolysin, γ-hemolysin and δ-hemolysin.It carries plv and tst virulence genes and is of high virulence .