2.Pyrosequencing technique for rapid identification of bacterial pathogens from body fluid specimens
Xuyi REN ; Jie LIN ; Runliu YU ; Jing ZHANG ; Jiangfeng Lü
Chinese Journal of Clinical Infectious Diseases 2011;04(6):333-338
Objective To develop a method for rapid and accurate detection and identification of bacterial pathogens directly from body fluid specimens and to evaluate its application in clinical laboratory.Methods Bacteria DNA was extracted from 205 body fluid specimens with column-based kit,and the high variable V1 and V3 regions of bacterial 16S rRNA gene were amplified with broad-range primers.Amplicons were analyzed by pyrosequencing and the generated sequences were searched in the bacterial identification database.Traditional culture-biochemical method was also used for these specimens and the results were taken as the golden standard.SPSS 11.0 was used to calculate the sensitivity,specificity,false positive/negative rate,positive/negative predictive value and positive/negative likelihood rate of pyrosequencing method.Results The positive rate of bacteria culture was 39.5% (81/205),among which 71 were infected with single bacterium,and 10 were infected with two species of bacteria.Compared with the culture identification results,pyrosequencing had a 100.0% (71/71) concordance when applied to detect and identify bacterial pathogens from specimens with single specie bacterium infected.To specimens with two species bacteria infected,7 out of 10 specimens were in concordance with the culture identification results.Besides,pyrosequencing detected 10 positive specimens and identified bacterial pathogens infected in the 124 culture-negative specimens.Taken bacteria culture as the standard method,the sensitivity of pyrosequencing for identifying bacterial pathogen in body fluid was 100.0%,and with a specificity of 91.9%,the false positive rate was 8.1%,the false negative rate was 0.0%,the positive predictive value was 89.0%,the negative predictive value was 100.0%,and the positive and the negative likelihood rate were 12.4 and 0,respectively.Conclusion Pyrosequencing can be used to detect and identify bacterial pathogens directly from body fluid specimens with the advantages of rapidity,high sensitivity,high accuracy and high throughput.
3.Enhancers on the transmembrane transport of chlorogenic acid.
Jing REN ; Shengqi DENG ; Xuehua JIANG ; Lingling WANG ; Yu XIAO
Acta Pharmaceutica Sinica 2014;49(2):252-5
To investigate the influence of the difference enhancers on the transport mechanism of chlorogenic acid (CGA) across Caco-2 cells model, a RP-HPLC method was adopted to detect the concentrations of CGA. At the concentrations of 20 to 80 microg x mL(-1), the difference of absorption rate constants (K(a)) was not statistically significant. At the concentrations of 40 and 20 microg x mL(-1), the ratios of apparent permeability coefficients (P(app)) of the apical to basolateral and the basolateral to apical were 1.14 and 1.18, respectively. With the effect of enhancers K(a) and P(app) increased, the absorption half-life (T1/2) decreased. CGA passed through the Caco-2 cell membrane mainly by passive transport. It showed that monocarboxylic acid transporter (MCT) could be involved in the across membrane transport process of CGA. Borneol had no effect on the cell membrane transport processes. The order of increasing absorption of CGA caused by the enhancers was sodium lauryl sulphate > sodium taurocholate > carbomer.
4.Removal of injected polyacrylamide hydrogel and repair of deformity after breast augmentation
Yuping REN ; Hui HUA ; Jing YU ; Yi XU ; Yiping WU
Chinese Journal of Medical Aesthetics and Cosmetology 2014;20(6):409-412
Objective To explore surgical methods of removing polyacrylamide hydrogel (PAHG) and the right time of repairing the deformity of breast after removing PAHG.Methods We operated with endoscopy to remove PAHG through the lower mammary areolar incision.According to injection influence,we made the dicision whether to put the silicon gel prosthesis (hereafter referred to as prosthesis) simultaneously,and to fix the porsthesis,and to rebuild the imframammary fold of breast using biological repair membrane (hereafter referred to as membrane).Results In all 46 patients,39 patients' injection and their envelope were removed entirely.7 patients left part of the envelope because of its thin and wide characters.14 of them accepted prosthesis augmentation mammaplasty simultaneously and 5 of these accepted membrane repair.4 patients received prosthesis augmentation in stage Ⅱ.All patients' incision were primary healing.The incision scars were not obvious.1 patient with breast cancer suffered breast excision; 1 patient who received prosthesis and membrane simultaneously appeared prosthesis displacement after 3 months and fixed again.1 patient who received membrane appeared hydrops in residual cavity,and the membrane was removed finally.Conclusions This method with endoscopy through mammary areolae is necessary for cleaning PAHG entirely.We can use prosthesis to repair the deformity of breast after removing PAHG,and if necessary use membrane to fix the implant and rebuild the inframammary fold of the breast.
5.Rapid identification of low activity of depleted uranium using portable γ-spectrometer
Tao YU ; Jing LIU ; Jiong REN ; Yuhui HAO ; Rong LI
Chinese Journal of Radiological Medicine and Protection 2015;35(3):217-221
Objective To explore the feasibility of using portable γ spectrometer to rapidly identify the low activity of depleted uranium and the underlying conditions.Methods Firstly,high purity germanium (Ge) γ spectrometer was used to analyze energy spectrum of DU samples (5 g) and calculate nuclide percentage of 235U in an attempt to ascertain the properties of these DU samples.The portable γ spectrometer was used to provide the evidences for identification of DU samples.Secondly,portable γ spectrometer was also used to identify DU samples of same group.These samples contain 1 g DU powder and 0-5 g environmental clay powder,which were sealed with double layer pocket,and then detected with a distance of 1-5 cm during the longest detection time of 10 min.According to the detection of nuclide activity of 238U and 235U in the samples and the subsequent calculation of specific activity,the nuclide percentage composition was calculated and the existence of DU was confirmed if this value of 235U was less than 0.718%.Results The activity of 238U was detected using portable γ spectrometer under all test conditions,while the activity of 235U was detectable only under certain test conditions (MA ≥ 1 g,DN ≤ 1 cm).Under the condition that the 238U and 235U was both detected,the nuclide percentage of 235U was all less than 0.718%,which suggested that the DU was confirmed.Conclusions The energy spectrum of low activity of DU and the type of nuclide could rapidly be identified and evaluated by using portable γ spectrometer.This is same as the conclusions obtained with high purity Ge γ spectrometer,α spectrometer and ICP-MS.
6.Pre-hepatic portal hypertension in a child.
Jian-hong SUN ; Li-hong REN ; Yu-jing ZHANG
Chinese Journal of Pediatrics 2006;44(12):955-955
7.Changes of Hemorheology of Children with Essential Hypertension
jing, TAO ; yu-chuan, WANG ; ren-bing, ZHAO
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To explore the hemorheological changes in children with essential hyepertension,and explore their clinical significance.Methods The hemorheological parameters,blood lipid,blood glucose in 37 children with essential hyepertension [19 boys and 18 girls,median age(12.49?3.20) years] were measured and analyzed.The parameters in 30 healthy children were measured as controls.SPSS 11.5 software was used to analyze the data.Results The whole blood viscosity under high,middle and low shear rate,plasma viscosity,hematocrit,rigidity index of erythrocyte,platelet aggregation M were significantly higher in hypertension group than those in healthy control group(Pa0.05).Conclusions There have been developed significant hemorheological changs in children with essential hypertension.The hemorheological changes might play an important role in pathogenesis of childhood hypertension.
8.Activation of STAT3 promoting immunosuppressive effect on T cells in MDSCs of breast cancer
Yue WANG ; Xiubao REN ; Hui LI ; Shui CAO ; Baozhu REN ; Wenwen YU ; Peng ZHANG ; Jing QI ; Jinpu YU
Chinese Journal of Clinical Oncology 2013;(17):1016-1019
Objective:To explore the status of STAT3 phosphorylation in myeloid-derived suppressor cells (MDSCs) of breast cancer and its function in the immunosuppressive effect of MDSCs on proliferation and cytokine secretion of T cells. Methods:CCD33+cells were isolated from healthy umbilical cord, blood-derived, peripheral blood mononuclear cells and were co-cultured with breast cancer cell line MDA-MB-231 in vitro using Transwell plates to induce MDSCs. The untreated CD33+cells were used as con-trols. Idoxuridine (IDO) suppressor expression and STAT3 phosphorylation were examined using Western blot assay. The proliferation and cytokine secretion of T cells, which were co-cultured with MDSCs, were determined by methyl thiazol tetrazolium assay and en-zyme-linked immunosorbent assay. 1-MT and JSI-124 were used to investigate the function of IDO and pSTAT3 in MDSC-mediated T cell immunosuppression. Results:The protein levels of IDO and pSTAT3 in MDSCs were significantly upregulated. MDSCs obviously suppressed T-cell proliferation, which was reversed by 1-MT or JSI-124 (P<0.05). MDSCs could promote TGF-βand IL-10 secretions, but could also remarkably inhibit IFN-γsecretion (P<0.05). After incubation with 1-MT or JSI-124, the increase in TGF-βand IL-10, as well as the decrease in IFN-γ, was significantly reversed. Conclusion:The upregulated pSTAT3 induced the IDO increase in MDSCs. JSI-124 can block MDSC-mediated immunosuppressive effect on T cells in breast cancer.
9.Effect of RetroNectin on the Proliferation of CIK Cells and the Possible Mechanisms
Ying HAN ; Jinpu YU ; Hui LI ; Shui CAO ; Baozhu REN ; Jing QI ; Xiumei AN ; Naining ZHANG ; Xiubao REN ; Xishan HAO
Chinese Journal of Clinical Oncology 2010;37(2):71-75
Objective: To investigate the effect of RetroNectin on CIKs cells and the related mechanisms. Methods: Peripheral blood mononuclear cells (PBMC) were collected from patients and divided into two groups: group Ⅰ and group Ⅱ. Samples in group Ⅰ were seeded into culture flask precoated with RetreNec-tin and CD3mAb to induce CIKs. While samples in group Ⅱ were seeded into common culture flask. The pro-liferation of CIKs was detected by cytometric analysis. The cytotoxic activity of CIKs was determined by LDH assays. The phenotype changes and cell cycle of CIKs were identified by flow cytometry. The apoptosis of cells was detected by Annexin V/PI. Western blot was employed to detect the level of protein Vav1. The CD49d and CD49e were blocked by anti-CD49d and anti-CD49e and the proliferation of cells was tested by cytometric analysis after the blockage. The phenotype changes of cells were identified by flow cytometry after the blockage. Results: RetroNectin enhanced the proliferation of CIKs (P<0.05). Flow cytometric analysis showed that RetroNectin significantly increased the number of CD25+ T cells (P<0.05). RN-CIK was more ac-tive than CIK in killing HCT-8 cell lines in vitro (P<0.05). RetroNectin could block the CIKs at G_1 phase (P<0.05) and resist apoptosis. There was no significant difference in the proliferation between the two groups af-ter the blockage with CD49d and CD49e (P>0.05). The expression of protein Vavl was associated with CD25+T cells. Conclusion: RetroNectin enhances the proliferation of CIKs by influencing the cell cycle, resist-ing apoptosis possibly through the site of CD49d and CD49e, and inducing T cell activation as the second sig-naling through Vav1.
10.Multidetector CT and magnetic resonance imaging features of solitary fibrous tumors in the pelvis and the relevant pathologic basis changes.
Xueming LI ; Jing REN ; Peng ZHOU ; Ying CAO ; Zhuzhong CHENG ; Jianqun YU ; Guohui XU
Journal of Biomedical Engineering 2015;32(1):157-162
In order to investigate the features of multidetector CT (MDCT) and magnetic resonance imaging (MRI) as well as the corresponding pathogic basis of solitary fibrous tumor (SFT) in the pelvis, we collected the clinical data of 13 patients with pathologically confirmed SFT in pelvis, and retrospectively reviewed the MDCT and MRI appearances. Of these enrolled patients, 6 received MDCT scans, 5 underwent MRI scans, and 2 underwent both MDCT and MRI examinations. Shown on the MDCT and MRI, the maximum diameters of the masses ranged from 4.0 to 25.2 cm (averaged 11.8 cm). Six masses were lobulated, and seven were round or oval. In addition, all masses were well-defined and displaced the adjacent structures to some degrees. On the computed tomography, all masses were of isodensity on unenhanced scans in general, among which five masses were demonstrated with hypodense areas. On the MRI T1-weighted image, all lesions were isointense, of which patchy hypointense areas were detected in 3 cases and radial hypointense areas were in 3 cases, and the other one was presented with homogenous intensity. On T2-weighted images, most of the lesions were mixed hyperintense, of which 3 cases were of heterogenous hyperintesity, radial hypointense areas were detected in 3 patients, and the other one was homogenously intense. On enhanced computed tomography and MRI, large supplying vessels were found in 4 cases; 12 cases showed moderate to conspicuous enhancement, and the other one was presented with mild homogenous enhancement. Of the patients with moderate to conspicuous enhancement, patchy areas of non-enhancement were detected in 7 cases, radial areas of progressive enhancement were detected in 3 cases, and the remained 2 cases showed homogenous enhancement. On pathology, the radial area presented as progressive enhancement was fibrosis. During the follow-ups after surgery, 2 patients had local recurrence and 1 had metastasis to liver. In conclusion, the SFT in the pelvis are commonly presented as a large solid, well-defined and hypervascular mass with necrosis or cystic changes at some extents together with the displacement of adjacent structures. The radial area with hypointensity on T2-weighted image and with progressive enhancement on enhanced magnetic resonance imaging is an important feature of SFT, which can be helpful for the diagnosis of this mass.
Humans
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Magnetic Resonance Imaging
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Neoplasm Recurrence, Local
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Pelvis
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pathology
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Retrospective Studies
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Solitary Fibrous Tumors
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diagnosis
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pathology
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Tomography, X-Ray Computed