1.The expression and clinical significance of serum follistatin-like protein 1 in patients with systemic lupus erythematosus
Lisha MO ; Qianghua WEI ; Dawei LI ; Yuli JIN ; Hongwei FU ; Liming QIAN
Chinese Journal of Rheumatology 2012;16(1):38-41
ObjectiveTo detect the serum level of follistatin-like protein 1 (FSTL1) in patients with systemic lupus erythematosus (SLE) and its expression in renal biopsy tissues in lupus nephritis (LN) patients as well as its clinical significance were analyzed.MethodsThe serum concentration of FSTL1 in 54 SLE patients and 27 healthy controls was measured with enzyme-linked immunosorbent assay (ELISA).The distribution of FSTL1 in renal biopsy tissues was stained by immune-histochemical method.Mann-Whitney U test,t test,X2test and Pearson test were selected to compare the changes and data analysis.ResultsThe serum FSTLI level was significantly higher in SLE patients(26±21) μg/L than those of healthy controls ( 12± 14) μg/L (P<0.01).The level of serum FSTL1 was significantly higher in SLE patients with hypertension than in patients without hypertension.The serum FSTL1 level had statistically significant changes between SLE patients with disease duration ≥ 5 years and <5 years.The level of serum FSTL1 correlated positively with SLEDAI score (r=0.319,P=0.022),age (r=0.700,P<0.01),disease duration (r=0.513,P<0.01),complement C4 level (r=0.443,P=0.004),and total serum cholesterol level (r=0.460,P=0.001 ).FSTL1 correlated inversely with platelet count (r=-0.422,P =0.001 ),anti-dsDNA antibody levels (r=-0.276,P=0.046).FSTL1 expression was evident in the cytoplasm of epithelial cells of kidney tubules.ConclusionThe level of serum FSTL1 is significantly increased in SLE patients.FSTL1 concentration correlats positively with disease activity.These data indicate that FSTL1 may play a role in the pathogenesis of SLE.
2.Preparation and immunological activity in vitro of human prostatic carcinoma-targeted liposome microbubbles
Lisha ZHUO ; Rui LI ; Yanli GUO ; Xing HUA ; Yun HE ; Zheng LIU ; Chixu FU
Journal of Third Military Medical University 2003;0(09):-
Objective To prepare human prostatic carcinoma-targeted ultrasound contrast agent and assess its targeted ability in vitro. Methods Human prostatic carcinoma-specific polyclonal antibody PSM(C-15) was attached to the surface of self-made liposome microbubbles by electrostatic attraction to prepare targeted microbubbles. Immunofluorescent staining assay was used to identify the combination of PSM(C-15) with liposome microbubbles and the targeted microbubbles were added to prostatic-carcinoma cells and then observed under the light and fluorescence microscope to evaluate the targeting ability of the targeted liposome microbubbles with prostatic carcinoma cells in vitro, while the common liposome microbubbles were controls. Results Targeted microbubbles were positive in immunofluorescent straining assay. In vitro study of the targeting ability showed the targeted microbubbles could actively adhere to LNCaP cell. While the control was negative. Conclusion The targeted liposome contrast agent with highly specific biological activity was prepared successfully. The contrast agent could bind to human prostatic carcinoma cells specially and effectively in vitro.
3.Synthesis of hyaluronic acid coated magnetic nanoparticles for highly efficient tumor CD44 cell labeling and in vitro study
Ruimeng YANG ; Nannan LI ; Liming ZHANG ; Li WANG ; Chaoping FU ; Danping HUANG ; Xinhua WEI ; Lisha LAI ; Xinqing JIANG
Chinese Journal of Radiology 2014;(5):363-368
Objective To explore a promising system for tumor CD 44 receptor-targeted imaging and to investigate their physic-chemical properties and targeting effect on CD 44 abundant cancer cells in vitro.Methods The superparamagnetic iron oxide ( SPIO) nanoparticles were prepared by a coprecipitation in alkaline media starting from a mixed of the ferrous and ferric solution.And then the surface of the SPIO nanoparticles were modified with APTMS by a reaction with the hydroxyl groups.Finally, the hyaluronan-modified SPIO ( SPIO-HA) nanoparticles were prepared.Control and experimental groups were established after adding SPIO or SPIO-HA as agents respectively.Transmission electron microscopy ( TEM) and particle size analyzer were used to measure these nanoparticle sizes and the hydrodynamic diameters.Thermogravimetric analysis ( TGA) was carried out to evaluate the HA-content on the surface of SPIO-HA.The MRI T2 ralaxivities (1/T2 ) of the two groups at different Fe concentrations (0.09, 0.18, 0.27, 0.36, 0.45 mmol/L ) were measured on a 3.0T MR system.HepG2 cells and HL7702 cells were used for assessment of cells viability by methyl thiazolyl tetrazolium ( MTT ) assay.Prussian blue staining , immunoassay fluorescence image and flow cytometry were carried out to determine the targeted cellular uptake of SPIO-HA nanoparticles.MRI were performed to show the MR T 2 value changes after incubating with HepG2 cancer cells by using T 2 WI sequences at a clinical 3.0 T MR system.One-way analysis of variance was performed to determine significant changes in MR T 2 values of blank control , SPIO-HA and SPIO groups.Results The SPIO-HA and SPIO NPs were fairly homogeneous with an average core size of 18.2 and 22.4 nm, hydrodynamic diameter of 91.1 and 103.2 nm, Zeta potential of (-45.00 ±0.86) mV and (-18.50 ±0.73) mV, and magnetic relaxivity of 0.212 ×106 M-1 · s-1 and 0.191 ×106 M-1 · s-1.Based on the TGA data , HA accounted for 24%weight of each SPIO-HA.The internalization of the SPIO-HA was confirmed by prussian blue staining , while the cells showed no obvious blue stains with SPIO , incubation of SPIO-HA with tumor cells led to blue color inside the cells.After that, we examined cancer cell binding of FITC-SPIO-HA by immunoassay fluorescence image and flow cytometry.The green fluorescence resulting from FITC-SPIO-HA was observed inside the cells in both the cytoplasm and the plasmalemma.Tumor cells treated with SPIO-HA exhibited higher fluorescence signals with 7.97-fold enhancement observed for HepG 2 cells over control particles.In vitro MR, mean T2 values of blank control , SPIO and SPIO-HA groups were ( 115.20 ±0.36 ), ( 115.07 ±0.81 ) and ( 21.67 ±0.21 ) ms, respectively.There was significant difference among those three groups (F=31 703.339,P<0.01), MR T2 values of HepG2 cells treated with the SPIO-HA NPs were lower than blank and SPIO group.In comparison, SPIO did not generate any MRI signal changes compared with blank group.Conclusion The tumor CD44 receptor-targeted MR molecular probe SPIO-HA had a good physic-chemical property and well targeted HepG2 cells.
4.Genomic characterization and cluster analysis of Carbapenem-resistant Klebsiella pneumoniae
Lijuan LI ; Ziyang YUAN ; Lu ZHANG ; Rentang DENG ; Lisha LAI ; Wencai HUANG ; Wenjin FU
Chinese Journal of Preventive Medicine 2024;58(9):1372-1378
To investigate the genomic features and perform cluster analysis of Carbapenem-resistant Klebsiella pneumoniae (CRKP) to provide an experimental basis for guiding the prevention and treatment of CRKP infections.A retrospective case-cohort study was conducted on 19 non-redundant CRKP strains isolated from the Tenth Affiliated Hospital of Southern Medical University between January and June 2023. Whole genome sequencing (WGS) and multilocus sequence typing (MLST) were performed to compare genomic features and analyze the resistance genes and homology of the strains.The results showed that the 19 CRKP strains were isolated from 8 different clinical departments, mainly from respiratory specimens. The whole genome sequencing revealed that the genomic lengths of CRKP ranged from 4.90 to 5.85 Mbp, with contigs N50 values>20 kb for each genome. The median overall GC content was 57.0% (50.4%-57.1%). Comparative genomic analysis identified three regions with high genomic variability. WGS detected 32 resistance genes across 11 categories. All 19 strains carried carbapenem resistance genes ( blaKPC-2 and blaOXA-48), blaTEM-1B extended-spectrum β-lactamase resistance genes, qnrS1 quinolone resistance gene, and fosA fosfomycin resistance gene, with each strain carrying only one carbapenemase gene. The detection rate of blaKPC-2 was 94.7% (18/19). MLST identified three sequence types: ST11, ST437 and ST147, with ST11 being predominant (89.5%, 17/19). Clustering analysis based on acquired resistance genes revealed three clonal transmission patterns among strains 72 and 90, and strains 88, 84, 66 and 79.In conclusion, CRKP strains carry multiple resistance genes, and clustering analysis indicating that nosocomial clonal transmission is closely related to acquired resistance genes. The ST11- blaKPC-2 type strain is the predominant clone. Strengthened surveillance and effective control strategies are necessary to reduce nosocomial transmission of CRKP.
5.Rapid detection of the bacterial drug susceptibility testing based on AIE technology
Lisha LAI ; Rentang DENG ; Lu ZHANG ; Yubang JIE ; Lingping XIE ; Zhihong HUANG ; Liming YIN ; Dujuan WANG ; Lijuan LI ; Junfa XU ; Lanfen PENG ; Wenjin FU
Chinese Journal of Laboratory Medicine 2023;46(11):1186-1192
Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.
6.Genomic characterization and cluster analysis of Carbapenem-resistant Klebsiella pneumoniae
Lijuan LI ; Ziyang YUAN ; Lu ZHANG ; Rentang DENG ; Lisha LAI ; Wencai HUANG ; Wenjin FU
Chinese Journal of Preventive Medicine 2024;58(9):1372-1378
To investigate the genomic features and perform cluster analysis of Carbapenem-resistant Klebsiella pneumoniae (CRKP) to provide an experimental basis for guiding the prevention and treatment of CRKP infections.A retrospective case-cohort study was conducted on 19 non-redundant CRKP strains isolated from the Tenth Affiliated Hospital of Southern Medical University between January and June 2023. Whole genome sequencing (WGS) and multilocus sequence typing (MLST) were performed to compare genomic features and analyze the resistance genes and homology of the strains.The results showed that the 19 CRKP strains were isolated from 8 different clinical departments, mainly from respiratory specimens. The whole genome sequencing revealed that the genomic lengths of CRKP ranged from 4.90 to 5.85 Mbp, with contigs N50 values>20 kb for each genome. The median overall GC content was 57.0% (50.4%-57.1%). Comparative genomic analysis identified three regions with high genomic variability. WGS detected 32 resistance genes across 11 categories. All 19 strains carried carbapenem resistance genes ( blaKPC-2 and blaOXA-48), blaTEM-1B extended-spectrum β-lactamase resistance genes, qnrS1 quinolone resistance gene, and fosA fosfomycin resistance gene, with each strain carrying only one carbapenemase gene. The detection rate of blaKPC-2 was 94.7% (18/19). MLST identified three sequence types: ST11, ST437 and ST147, with ST11 being predominant (89.5%, 17/19). Clustering analysis based on acquired resistance genes revealed three clonal transmission patterns among strains 72 and 90, and strains 88, 84, 66 and 79.In conclusion, CRKP strains carry multiple resistance genes, and clustering analysis indicating that nosocomial clonal transmission is closely related to acquired resistance genes. The ST11- blaKPC-2 type strain is the predominant clone. Strengthened surveillance and effective control strategies are necessary to reduce nosocomial transmission of CRKP.
7.The role of erlotinib in the acute lung injury and the expression of surfactant protein A
Huan TAO ; Younian XU ; Lisha FU ; Huimin XIA ; Shanglong YAO ; Shihai ZHANG
Chinese Journal of Emergency Medicine 2018;27(8):881-886
Objective To investigate the role of erlotinib in the expression of surfactant protein A (SP-A) in LPS-induced acute lung injury (ALI) of mice model.Methods C57BL/6 mice were randomly (random number)divided into control group (n=6),ER group (n=6),LPS group (n=6),and ER+LPS group (n=6).In the LPS group,2 mg/kg LPS was instilled into trachea of mice to induce lung injury.In control group,normal saline was instilled into trachea of mice instead.In the ER+LPS group and ER group,100 mg/kg of edotinib was instilled into stomach of mice,and one hour later.2 mg/kg LPS was instilled into trachea of mice in ER+PLS group to induce lung injury.Twenty-four hours later,bronchoalveolar lavage fluid (BALF) and lung tissue of mice in four groups were collected.HE staining were used for evaluating pathological changes of lung injury.Lung wet/dry weight ratio,protein concentrations and total cell numbers in the BALF were measured to determine the degree of pulmonary edema.Immunohistochemical staining and Western Blot were used for testing the protein expression of SP-A,Data of multiple groups were analyzed by one way variance (ANOVA) and inter-group comparisons were made by the least significant difference (LSD) tests.Results There was no significant difference in lung injury score (LIS) between control group (0.056±0.008) and ER (0.064±0.037) group,The LIS in LPS group (0.846-±0.047) was higher than that in control group,however the LIS in ER+LPS group (0.279±0.020) was significant lower than that in LPS group (P < 0.05).Lung wet/dry weight,SP-A concentration and total cell numbers in the bronchoalveolar lavage fluid revealed that the degree of pulmonary edema in LPS group was higher than that in control group,and this pulmonary edema was reversed by erlotinib treatment.Immunohistochemical staining and Western blot showed that the expression of SP-A in LPS group was decreased compared with control group,but it was recovered after erlotinib treatment (P < 0.05).Conclusions Erlotinib could protect the LPS-induced ALI,and it may be related to the regulation of SP-A.
8.Palm vein recognition based on end-to-end convolutional neural network.
Dongyang DU ; Lijun LU ; Ruiyang FU ; Lisha YUAN ; Wufan CHEN ; Yaqin LIU
Journal of Southern Medical University 2019;39(2):207-214
We propose a novel palm-vein recognition model based on the end-to-end convolutional neural network. In this model, the convolutional layer and the pooling layer were alternately connected to extract the image features, and the categorical attribute was estimated simultaneously via the neural network classifier. The classification error was minimized via the mini-batch stochastic gradient descent algorithm with momentum to optimize the feature descriptor along with the direction of the gradient descent. Four strategies including data augmentation, batch normalization, dropout, and L2 parameter regularization were applied in the model to reduce the generalization error. The experimental results showed that for classifying 500 subjects form PolyU database and a self-established database, this model achieved identification rates of 99.90% and 98.05%, respectively, with an identification time for a single sample less than 9 ms. The proposed approach, as compared with the traditional method, could improve the accuracy of palm vein recognition in clincal applications and provides a new approach to palm vein recognition.
Algorithms
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Databases, Factual
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Hand
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blood supply
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diagnostic imaging
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Humans
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Neural Networks (Computer)
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Veins
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diagnostic imaging
9. Rapid detection of CYP2C9, CYP2C19,CYP4F2,VKORC1 and ABCB1 gene polymorphisms by liquid phase chip technology
Hongli XU ; Rentang DENG ; Meilian CHEN ; Zaixin CHEN ; Zhihong HUANG ; Bo SITU ; Guixing KONG ; Lisha LAI ; Lei ZHENG ; Wenjin FU
Chinese Journal of Laboratory Medicine 2019;42(12):1042-1050
Objective:
To establish a method for simultaneous and rapid detecting of the polymorphisms in Cytochrome P450 2C9 (
10. Metabolomics investigation on antiobesity effects of Corydalis bungeana on high-fat high-sugar diet-induced obese rats
Minghai FU ; LiSha A. ; Sungbo CHO ; Minghai FU ; Terigele BAO ; Hongzhen YU ; HuiFang LI ; Genna BA ; Sungbo CHO
Chinese Herbal Medicines 2022;14(3):414-421
Objective: Corydalis bungeana (CB) is a well-used medicinal herb in Mongolian folk medicine and has been traditionally applied as an antiobesity agent. However, the evidence-based pharmacological effects of CB and its specific metabolic alterations in the obese model are not entirely understood. This study aimed to utilize untargeted metabolomic techniques to identify biomarkers and gain mechanistic insight into the serum metabolite alterations associated with weight loss and lipid metabolism in obese rats. Methods: A high-fat high-sugar (HFHS) diet was used to induce obese models in rats. CB extract was orally gavaged at 0.18, 0.9 and 1.8 g/kg doses for six weeks, and feed intake, body weight, fat pad weight, and blood indexes were measured. Blood serum metabolites were evaluated by gas chromatography/quadrupole time-of-flight tandem mass spectrometry (GC-TOF/MS). Results: The results showed that compared with the obese group, the administration of CB extract caused significant decreases in body weight (P < 0.05), feed intake, Lee's index, and perirenal, mesenteric, epididymal fat weight. CB extract also reduced blood triglyceride and total cholesterol levels (P < 0.05) of obese rats. Metabolomic findings showed that nine differential metabolites, including pyruvic acid, D-glucuronic acid, malic acid, dimethylglycine, oxoglutaric acid, pantothenic acid, sorbitol acid, fumaric acid and glucose 6-phosphate were identified under CB treatment and altered metabolic pathways such as TCA cycle, pantothenate and CoA biosynthesis, and glycolysis/gluconeogenesis. Conclusion: This study demonstrated weight loss and lipid lowering effects of CB on HFHS diet-induced obese rats and identified nine metabolites as potential biomarkers for evaluating the favorable therapeutic mechanism of CB via regulation of lipid and glucose metabolism.