1.Study on bacteria contamination during the collection, processing and storage of umbilical cord blood
Leling ZHANG ; Lixia MA ; Sulan WANG ; Xindang WANG ; Fu LI ; Lehai ZHANG ; Xingli LIU ; Baijun SHEN
International Journal of Pediatrics 2012;39(2):212-214
ObjectiveTo evaluate bacteria contamination during collection,processing and storage of cord blood to gain insight into contamination mechanism and direct prevention.MethodsFresh cord blood was separated by hydroxyethyl starch (HES) to harvest nucleated cells.The bacteria contamination was tested by culturing 10 ml plasma-red cells with BacT/ALERT 3D-480 automatic blood culture system.Total 87 positive samples were further identified for bacteria species.Ninety six cord blood nucleated cells concentrate with bacteria positive stored in liquid nitrogen(LN2) for 6-7 years were thawed at 37 C and re-cultured for bacteria analysis.ResultsWe collected 19 062 umbilical cord blood.Among them,336 was bacteria positive ( contamination rate 1.8 % ).Eighty-seven positive samples were further investigated with facultative bacteria 58 (66.7 % ),aerobic 38(43.7% ) and anaerobic 17( 19.5% ),Gram- negative accounted for 68% while positive 32%.The most frequent bacteria were Escherichia coli ( 25.3% ),Streptacoccus intermediate ( 14.9% ) and Chromobacteria violaceum(9.2% ).Ninety-six nucleated cells concentrate with bacteria positive were cryopreserved at liquid nitrogen for researching.Of them,83 samples( 86% ) showed positive of bacteria culture after deep-low temperature storage for 6-7 years.ConclusionsBacteria contamination rate of the cord blood collection,processing and storage in 2000 ~ 2007 was 1.8%.Stored in liquid nitrogen for 6-7 years,the viability of bacteria was 86%.The aseptic procedures of cord blood collection in delivery room should be intensified.The bacteria re-culture following thawing of cord blood cells is necessary before clinical transfusion.
2.SMN1 gene deletion analysis using mid-trimester amniotic fluid cells by real-time PCR
Heyu HU ; Xingli FU ; Jing YU ; Bei ZHANG ; Zhenhua TANG ; Huifen CHEN ; Li ZHANG ; Bo GONG
Chinese Journal of Laboratory Medicine 2016;39(6):418-422
Objectives To investigate the prenatal diagnosis method of spinal muscular atrophy with amniotic fluid sample.Methods Totally 1 064 amniotic fluid samples from mid-trimester pregnant women were enrolled during January 2015 and January 2016 in 4 hospitals.Genetic analysis was performed for detecting potential contamination of maternal tissue by a genetic technique based on short tandem repeat ( STR) markers.Deletion of SMN1 gene was detected in 1 062 uncontaminated amniotic fluid samples by real-time PCR and multiplex ligation-dependent probe amplification ( MLPA) respectively.Results Two contaminated amniotic fluid samples were detected within 1 064 mid-trimester pregnant women by STR genotyping.The other 1 062 uncontaminated amniotic fluid samples were tested by real-time PCR.There were 37 samples with heterozygous deletion of Exon 7 of SMN1 gene ( 3.67%) , 34 samples with heterozygous deletion of Exon 8 of SMN1 gene (3.2%) and two samples with homozygous deletion of Exon 7 and Exon8 of SMN1 gene ( 0.19%) respectively , while other samples observed with no deletion of Exon 7 and Exon8 in SMN1 gene.Totally 41 samples with heterozygous or homozygous deletion of SMN 1 gene and 55 samples with undetected deletion of SMN 1 gene were confirmed by MLPA and the results showed 100%consistence with that of real-time PCR.Conclusions Both real-time PCR and MLPA are suitable for detecting the deletion of SMN 1 gene with amniotic fluid sample . Real-time PCR exhibits less sample requirement and time compared with MLPA .
3.Methodological study of quantitative detection of Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA
Zhen WANG ; Yuhua GONG ; Caidi QIAN ; Chunhong SUN ; Liping ZHOU ; Xingli FU ; Qixing SHAO
International Journal of Laboratory Medicine 2014;(21):2931-2933
Objective To establish a quantitative detection method for Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA detection system with double internal standards(IMC-PCR-ELISA) .Methods The immunomagnetic (Dynabeads? ) which could specifically capture Mycobacterium tuberculosis were prepared .According to Mtp40 and IS6110 gene sequence of Mycobacterium Tuberculosis ,2 pairs of primers(upstream primer was modified with Biotin at 5′end) ,2 same-length mutant fragments with PCR amplified fragments ,and 3 capture probes(modified with digoxigenin at 3′end) were designed .Myco-bacterium tuberculosis were captured by immunomagnetic ,then detected by PCR-ELISA with double internal standards .Results The IMC-PCR-ELISA method could yield quantitative results in about 4 h with a detection limit at 5 × 103 copies/mL .There was a fine linear relationship between the copies of Mtp40(IS6110)in fact and in the calculation through formula when the concentrations of low internal standards were 30-70 copies/mL and the concentrations of high internal standards were 8 000-12 000 copies/mL (r2 =0 .998) .No nonspecific amplification was observed .Conclusion A rapid and quantitative method for the detection of Myco-bacterium tuberculosis was established successfully .The IMC-PCR-ELISA method was rapid ,sensitive ,secific and quantitative .
4.Exploration on Permanent Staining Preparation of Tapeworm
Shengxia CHEN ; Liang WU ; Huijuan XU ; Lianyun SHUAI ; Xingli FU ; Jinbo QIU
Chinese Journal of Parasitology and Parasitic Diseases 2006;0(S1):-
Objective To make staining preparation of tapeworm for the purpose of teaching and scientific research.Methoeds Pregnant segmens were stained by Indian ink,and mature segments,cephalomeres and bladder worms were stained by carmine.Result After staining the characteristics of pregnant segments,mature segments,cephalomeres and bladder worms were observed obviously.For instance,the stem and branches linking both-side of uteri in pregnant segment and reproduction cavity projecting in one of edges in the center of the pregnant segment were clearly shown in ink colour after stained by Indian ink.The red colour was shown in the mature segment with deep-coloured male and female reproduction systems inside,separately,after stained by carmine.Conclusion Tapeworm preparation stained by Indian ink and carmine is useful and can be preserved permanently.
5.Application of diffusion tensor imaging FA value and ADC value in glioma grading:A Meta-analysis
Ruizhe HOU ; Yingjia ZHAO ; Yan LIU ; Qunbang CHEN ; Hong FU ; Jingyan CHEN ; Xingli ZHAO
Journal of Jilin University(Medicine Edition) 2017;43(2):311-316
Objective:To clarify the significance of the fractional anisotropy(FA) value and apparent diffusion coefficient(ADC) value of diffusion tensor imaging(DTI) applied in the glioma grading with Meta-analysis, and to provide a reference for selecting the treatment program and judging the prognosis of glioma.Methods:Computer search was performed in PubMed database, EMBASE database,CBM, CNKI, Wanfang database, and VIP database.The relevant neurosurgery magazines and neuroimaging magazines, the proceedings of academic meeting were retrieved by hand and the references of related reviews were retrospectively retrieved.Then the case-series studies according to inclusion and exclusion criteria were screened,and the qualities of included studies were evaluated.Meta-analysis was carried out by Review Manager 5.2 software.Results:Nine studies were included.The FA value of low-grade glioma(LGG) in tumor centre was lower than that of high-grade glioma(HGG) (χ2=11.94,df=8,P=0.15;I2=33%,Z=2.78,P=0.005;WMDFA=-0.02,95%CI:-0.03--0.01),and the ADC value of LGG in tumor centre was higher than that of HGG (χ2=5.26,df=4,P=0.26;I2=24%,Z=2.19,P=0.03;WMDADC=0.15,95%CI:0.02-0.28).Conclusion:When the FA value in tumor centre of the glioma is increasd,the ADC value is decreased,and the grade of glioma is increasd.The FA value and ADC value of DTI can help determine the grade of gliomas.
6.Pulmonary embolism complicated with abnormal spinal cord embolism due to lack of protein S activity: a case report
Xingli TAN ; Qingxi FU ; Liangjun JIANG ; Naiyong GAO
Chinese Journal of Neurology 2018;51(11):911-913
Spinal vascular disease is rare in clinic, especially in cases of spinal cord embolism caused by abnormal embolism. It is rare in clinic and has not been reported in China. We report a case of pulmonary embolism complicated with abnormal spinal cord embolism due to lack of protein S activity and analyze its clinical data in order to improve clinical understanding of the disease.
7.Overexpression of miR-155-5p Inhibits the Proliferation and Migration of IL-13-Induced Human Bronchial Smooth Muscle Cells by Suppressing TGF-β-Activated Kinase 1/MAP3K7-Binding Protein 2.
Yujia SHI ; Xingli FU ; Qi CAO ; Zhengdao MAO ; Yi CHEN ; Yun SUN ; Zhiguang LIU ; Qian ZHANG
Allergy, Asthma & Immunology Research 2018;10(3):260-267
PURPOSE: Molecular mechanisms leading to asthma is still ill-defined. Though the function of microRNAs (miRNAs) in asthma was previously reported, the involvement of miR-155 in important features of this disease remains unknown. The present study was designed to uncover the probable involvement of miR-155-5p in the proliferation and migration of IL-13-induced human bronchial smooth muscle cells (BSMCs) and the intrinsic regulatory mechanism. METHODS: The effects of different concentrations of IL-13 on the proliferation and migration of BSMCs as well as the expression of miR-155-5p and its predicted target transforming growth factor (TGF)-β-activated kinase 1/MAP3K7-binding protein 2 (TAB2) were investigated. The effects of miR-155-5p on the proliferation and migration of interleukin (IL)-13-induced BSMCs was determined in vitro using BSMCs transfected with miR-155 mimic/inhibitor and induced by a high concentration of IL-13. The quantitative real-time polymerase chain reaction (qRTPCR) was employed for determining the expression of miR-155-5p and TAB2. Western blotting was applied to analyze the expression of TAB2 at the protein level. Cell proliferation and migration were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Transwell assays, respectively. RESULTS: The proliferation and migration of BSMCs were dose-dependently increased with IL-13 treatment. Contrariwise, IL-13 dose-dependently inhibited the expression of miR-155-5p in BSMCs. Mechanistic studies showed that inhibition of miR-155-5p further promoted the stimulatory effects of IL-13, whereas overexpression of miR-155 significantly inhibited these effects. In silico studies and luciferase reporter assays indicated that TAB2 was a negatively regulated miR-155-5p target. CONCLUSIONS: These results suggested that miR-155-5p-inhibit the IL-13-induced proliferation and migration of BSMCs by targeting TAB2 and that the IL-13/miR-155/TAB2 pathway could serve as a therapeutic target for pulmonary diseases, especially asthma.
Asthma
;
Blotting, Western
;
Cell Proliferation
;
Computer Simulation
;
Humans*
;
In Vitro Techniques
;
Interleukin-13
;
Interleukins
;
Luciferases
;
Lung Diseases
;
MicroRNAs
;
Muscle, Smooth*
;
Myocytes, Smooth Muscle*
;
Phosphotransferases*
;
Real-Time Polymerase Chain Reaction
;
Transforming Growth Factors
8.Classification of tibia plateau fracture according to the "four-column and nine-segment"
Xiang YAO ; Yong XU ; Jishan YUAN ; Bin LYU ; Xingli FU ; Lei WANG ; Shengquan YANG ; Sheng MENG
Chinese Journal of Orthopaedic Trauma 2020;22(8):665-675
Objective:To characterize tibial plateau fractures using a computed-tomography-based "four-column and nine-segment" classification.Methods:A retrospective analysis was conducted of the 698 adult patients with tibial plateau fracture (704 knees) who had been admitted to Department of Orthopedics, The Affiliated People's Hospital of Jiangsu University from December 2007 to May 2018. They were 377 males and 321 females with an average age of 51.6 years. The left knee was affected in 371 cases (53.2%), the right knee in 321 cases (46.0%) and bilateral knees in 6 cases (0.9%). According to the differentiated morphological characteristics, the tibial plateau and proximal fibula were divided into 4 columns, which were subdivided into 9 segments. Tibial plateau injury index (TPII) was innovatively introduced to represent the extent of injury. Fracture mapping was retrospectively analyzed according to the "four-column and nine-segment" classification based on the CT imaging.Results:The rates of one-column, two-column, three-column and four-column injuries were 30.5% (215/704), 31.5% (222/704), 28.0% (197/704), and 9.9% (70/704), respectively. On average, 2.2 columns ± 1.0 columns and 3.6 segments ± 2.1 segments were affected in each case. The mean TPII was 5.7±3.0. The rates of mild, moderate and severe comminuted fractures were 50.0% (352/704), 37.5% (264/704), and 12.5% (88/704). The columns most frequently affected were the lateral column (572, 81.3%) and the intermedial column (524, 74.4%) while the less frequently involved ones the medial column (219, 31.1%) and the fibular column (218, 31.0%). The sequence of the segments affected was the posterolateral segment (465, 66.1%), the anterolateral segment (453, 64.3%), the posteromedian segment (379, 53.8%) and the tubercle segment (85, 12.1%).Conclusions:The novel "four-column and nine-segment" classification may be a beneficial system for clinical diagnosis, statistical analysis and prognostic judgment of tibial plateau fractures.
9.Design of Real-time Disease Warning System Based on Storm Technology.
Xingli YANG ; Yongquan FU ; Fangyu HU ; Liang LIANG ; Xingang LEI ; Jiwu ZHANG
Chinese Journal of Medical Instrumentation 2018;42(1):35-37
With the improvement of the quality of clinical diagnosis and treatment, the traditional scheduled "ward round" mode cannot meet the demands for real-time monitoring of acute and critically ill patients. This paper introduces the Storm, a real-time data stream processing technology and its application in the real time disease early warning system. By collecting the clinical data flow and calculating the MEWS scores in real time, the system can identify the potential deterioration of the disease, and promptly notify the medical staff. Score calculation results can be stored for further analysis and presentation as well.
Clinical Alarms
;
Critical Illness
;
Humans
10. Regulatory effect of metabolic glutamate receptor 1 on maltolate aluminum-induced synaptic plasticity in rats
Yaqin LI ; Baolong PAN ; Ye FU ; Liang LI ; Xingli XUE ; Huifang ZHANG ; Qiao NIU
China Occupational Medicine 2020;47(01):24-29
OBJECTIVE: To observe the effect of maltolate aluminum on synaptic plasticity in the hippocampus of rats and to explore the regulatory effect and mechanism of metabotropic glutamate receptor 1(mGluR1). METHODS: Specific pathogen free healthy adult male SD rats were randomly divided into control group, aluminum group, aluminum agonist group and aluminum antagonist group, 8 rats in each group. The rats in the control group received no treatment; the rats in aluminum group were injected with 5 μL 10 mmol/L maltolate aluminum solution into the lateral ventricle; the rats in aluminum agonists and aluminum antagonist group were injected with 3 μL 10 mmol/L maltolate aluminum solution plus 2 μL 0.1 μmol/L mGluR1 agonist or 2 μL 0.2 μmol/L mGluR1 antagonists into the lateral ventricle, respectively.Maltolate aluminum solution was injected every 2 days and continued for 10 days. After maltolate aluminum exposure, the amplitudes of long-term potentiation(LTP) in hippocampal CA1 region of rats were measured, and the relative expression levels of mRNA and protein of mGluR1, N-methyl-D-aspartate receptor(NMDAR1) and protein kinase C(PKC) in hippocampus tissue of rats were detected by real-time fluorescence quantitative polymerase chain reaction and Western blotting. RESULTS: The amplitude of LTP in hippocampal CA1 region in aluminum group and aluminum agonist group was lower than that in the control group and the aluminum antagonist group(P<0.05). Compared with the control group, the relative expression of mGluR1 mRNA and protein in the aluminum group increased, the relative expression of PKC and NMDAR1 mRNA and protein in the aluminum group decreased(P<0.05). Compared with the aluminum group, the relative expression of mGluR1 mRNA and protein in the aluminum agonist group increased, while the NMDAR1 mRNA decreased(P<0.05); the relative expression of mGluR1 mRNA and protein in the aluminum antagonist group decreased, while the NMDAR1 mRNA and protein increased(P<0.05). Compared with the aluminum agonist group, the relative expression of mGluR1 mRNA and protein decreased, while the NMDAR1 mRNA and protein increased in the aluminum antagonist group(P<0.05). The relative expression level of PKC mRNA and protein in aluminum agonist group and aluminum antagonist group was not statistically significant(P>0.05), and there was no statistical significance in these two groups compared with control group and aluminum group(P>0.05). CONCLUSION: Maltolate aluminum exposure can inhibit synaptic plasticity by inhibiting LTP in hippocampus of rats, and the mechanism may be related to the regulation of NMDAR1 expression by mGluR1.