Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 82
Filter
1.
Article in Chinese | WPRIM | ID: wpr-939707

ABSTRACT

OBJECTIVE@#To establish the technique that take the advantages of flow cytometry combined fluorescence in situ hybridization (Flow-FISH) to identify the Epstein-Barr virus(EBV) infected lymphocyte subtypies in patients' peripheral blood sample.@*METHODS@#Peripheral Blood monocyte from 9 patients with EBV infection enrolled at Children's Hospital in Chongqing Medical University were isolated by Ficoll-paque centrifugal separation. The expressions of EBER1, EBER2 in cell were detected by qRT-PCR. The surface markers of cell were detected by Flow cytometry after staining with their antibodies. The cell was treated Fix-Permeabilization Buffer before hybridization with fluorescent labeled probe at 37 ℃ overnight. The cell status, surface markers and targeted mRNA are detected by flow cytometry and fluorescence microscope.@*RESULTS@#It was optimized that the Fix-Permeabilization Buffer and recipe with 0.2% Tween-20 were picked out as providing a good cell integrity and high resolution of surface markers. Hybridization with 20% formamide and 7% dextran sulfate at 37 ℃ overnight is the optimal hybridization condition as a good hybridization effect, a detectable cell integrity and a high resolution of cell markers under flow cytometry detection. Finally, upon the established Flow-FISH method, lymphocyte subpopulations of the EBV+ cells from cell lines and blood samples of patients were identified successfully.@*CONCLUSION@#A Flow-FISH technology is established, which can be applied in the identification of EBV infected cell subtypes. This research provides a foundmental for its application in clinical test in EBV+ related proliferative diseases.


Subject(s)
Epstein-Barr Virus Infections , Flow Cytometry/methods , Herpesvirus 4, Human , Humans , In Situ Hybridization, Fluorescence/methods , Lymphocyte Subsets
2.
Article in Chinese | WPRIM | ID: wpr-934436

ABSTRACT

Objective:To explore the critical issues in the construction of management and supporting system of Investigator-Initiated trials (IIT) in pediatrics.Methods:Through summarizing related literature and considering the current status of pediatric clinical research, the critical issues in the construction of management and support system, for instance, the responsibility, training model, and performance evaluation in Europe and U. S. were evaluated, decision-making suggestions were put forward based on domestic pediatric IIT management system.Results:Besides IIT, clinical trials on children′s drugs are also supported by the pediatric clinical research management system in Europe and U. S.. The supporting service covers research consultation, ethical review, research design, trial implementation, patient education, risk control, and investigator training. The organizational structure and management system are relatively mature. Clinical trials are the majority of clinical research in children′s hospitals in China. Main issues identified in the construction of the management and supporting system include ethical review for pediatric clinical research, professional investigator training, multicenter cooperation scheme, performance assessment, and incentive strategies.Conclusions:Taking account into the current status of pediatric IIT in China, it is urgent to accelerate the training of pediatric investigators, set up standard IIT project management team, build the professional project management platform and Electronic Data Capture System, and promote the transformation of research outcomes. Finally, the whole process management of pediatric IIT will be developed to facilitate the development of pediatric medicine.

3.
Journal of Experimental Hematology ; (6): 1456-1461, 2021.
Article in Chinese | WPRIM | ID: wpr-922279

ABSTRACT

OBJECTIVE@#To investigate the effect of β-arrestin1 on the concentration of reactive oxygen species (ROS) in the mitochondria of acute T-lymphocytic leukemia (T-ALL) cells and its possible mechanisms.@*METHODS@#The stable T-ALL cell line with knocked down β-arrestin1 (Jurkat Siβ1) was constructed. Flow cytometry and probe assays were used to detect ROS content in cell and mitochondrial, respectively. The relationship between β-arrestin1 and microRNA was detected, analyzed and Q-PCR confirmed by microRNA microarray. The target genes of microRNA were predicated by miRbase software, identified by Western blot, and validated by Dual luciferase reporter gene.@*RESULTS@#Jurkat Siβ1 stable cell line was successfully constructed and it was found that ROS content was slightly reduced in Jurkat Siβ1 at the whole cell level, and the ROS content was also significantly reduced in mitochondria. MicroRNA microarray analysis revealed that multiple T-ALL related microRNAs showed differentially expressed, in which the expression of miR-652-5p was significantly increased in Jurkat Siβ1 (P2.0), and Q-PCR showed that miR-652-5p was nearly 5-fold up-regulated in Jurkat Siβ1. miRbase predicted that the P62 gene was the target gene of miR-652-5p which could regulates mitochondrial function. P62 protein showed highly expressed in stably knocked down miR-652-5p in Jurkat cells. Dual luciferase reporter gene assay confirms that P62 was the target gene of miR-652-5p.@*CONCLUSION@#β-arrestin1 can decreases the expression of miR-652-5p and deregulates the translational inhibition of P62 mRNA, thus to increase ROS content in mitochondria of T-ALL cells.


Subject(s)
Humans , MicroRNAs/genetics , Mitochondria , Oxygen , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma , RNA, Messenger , beta-Arrestin 1
4.
Article in Chinese | WPRIM | ID: wpr-885801

ABSTRACT

Objective:The hemolytic prediction model of the axial flow impeller blood pump is carried out by using a computational fluid dynamics(CFD) multiphase flow model.Methods:The hydrodynamic performance of the pump and the flow field in the pump, and the shear stress distribution are analyzed. A hemolytic prediction model based on the shear stress is built based on the calculation results. Hemolysis tests in vitro were performed 6 times with fresh bovine blood. At each time, the flow of the ventricular assist device(VAD) is 5 L/min and the outflow tract pressure is 100 mmHg(13.3kPa). According to the tests, the plasma free hemoglobin(FHB) content and the hematocrit(HCT) are measured every half hour. At the end of each experiment Normal Index of Hemolysis(NIH) is calculated.Results:The average of NIH is 0.0055 g/100L, almost identical with that obtained from the hemolytic prediction model.Conclusion:Multiphase flow model can be used for quantitative predictions of the hemolytic behavior of a VAD. This method can be applied in the selection stage of a blood pump.

5.
Article in Chinese | WPRIM | ID: wpr-871307

ABSTRACT

Objective:To investigate the causes of immune failure in the population with high vaccination rate of measles and rubella vaccine in Beijing by detecting the IgG antibody affinity in suspected cases of measles and rubella.Methods:Serum samples of 276 suspected cases of measles and rubella were tested for IgM and IgG antibodies by enzyme-linked immunosorbent assay (ELISA). The affinity of IgG antibody was detected, and the relative affinity index was calculated.Results:Among the 276 suspected cases, 104 were measles and 108 were rubella. Six measles cases had vaccination history and were caused by primary immunization failure ( n=3) and secondary immunization failure ( n=3). Twelve rubella cases had vaccination history and were due to primary immunization failure ( n=4) and secondary immunization failure ( n=8). Specific high-affinity antibodies were detected in nine measles cases and seven rubella cases without vaccination history, which indicated that these cases were reinfected. In the cases without measles or rubella, other pathogenic infections including mixed infections were detected, which were mainly caused by EB virus. Conclusions:Both primary and secondary immunization failure occurred in the population with immunization history. Reinfection was found in the patients who had not received vaccination against measles or rubella. Other pathogenic infections were existed among the cases without measles or rubella. Thus, misdiagnosis was responsible for the increased proportion of measles and rubella patients with immunization history in suspected cases in recent years. Full-course vaccination was conducive to produce high-affinity antibodies against measles and rubella. A supplementary vaccination campaign should be launched to consolidate the immune barrier against measles and rubella in key population or high-risk population, aiming to block the circulation of measles virus and achieve the goal of eliminating measles.

6.
Article in Chinese | WPRIM | ID: wpr-871296

ABSTRACT

Objective:To confirm the possible pathogen causing an outbreak of respiratory infectious disease in Beijing.Methods:Oropharyngeal swabs were collected from 14 cases with fever and detected by RT-PCR for respiratory viruses and bacteria. For specimens positive for adenoviruses, Fiber, Hexon and Penton gene fragments were amplified with specific primers and sequenced. BLAST and phylogenetic tree were used for sequence analysis.Results:All of the 14 specimens were adenovirus-positive. BLAST analysis of the sequences of Fiber, hexon and Penton genes showed that the 14 cases were all caused by adenovirus 3. The phylogenic tree analysis indicated that this adenovirus was closely related to an adenovirus of 3a51 genotype (GenBank No: KF268123) isolated in the USA in 2007.Conclusions:Human adenovirus genotype 3a51 caused this outbreak of respiratory infectious disease in Beijing.

7.
Chinese Journal of Endemiology ; (12): 791-795, 2020.
Article in Chinese | WPRIM | ID: wpr-866210

ABSTRACT

Objective:To analyze the gene mutation types and haematological characteristics of αβ compound thalassemia, non-delectable α-thalassemia and Hemoglobin H Disease (HbH disease) in Foshan.Methods:Using the method of retrospective analysis, we selected the population who had been tested for thalassemia gene in Foshan Second People's Hospital Affiliated to Southern Medical University from January 2011 to November 2019. Sysmex XT-5000 automatic hematology analyzer was used for routine blood analysis. α-, β- thalassemia genes were detected by PCR + diversion hybridization method.Results:A total of 4 563 people were tested, of which 1 829 were diagnosed as thalassaemia through genetic diagnosis. αβ compound thalassaemia was detected in 81 cases with a positive rate of 1.8%; non-delectable α-thalassemia was detected in 18 cases with a positive rate of 0.4%; HbH disease was detected in 23 cases with a positive rate of 0.5%. The most common genotypes of αβ compound thalassemia were -- SEA/αα\β41-42/βN (17.3%, 14/81), -α 3.7/αα\β41-42/βN (14.8%, 12/81), -- SEA/αα\β654/βN (11.1%, 9/81). The main manifestations of hematology were normal to mild anemia (93.8%, 76/81). Only β-thalassemia with double heterozygotes and α-thalassemia showed severe anemia. αα CS/αα\βN/βN genotypes were common in the local non delectable α-thalassemia (50.0%, 9/18), and the non delectable α-thalassemia was characterized by non-positive phenotype or typical small-cell hypochromatosis in hematology. The genotypes of local HbH patients were -α 3.7/-- SEA\βN/βN (65.2%, 15/23), and simple HbH manifested as moderate anemia (87.0%, 20/23). Patients with HbH disease and β-thalassemia had normal or mild anemia (13.0%, 3/23). Conclusions:The genotypes of αβ compound thalassemia in Foshan area are diverse and complex, and hematology mainly manifests as mild anemia or normal. Non-delectable α-thalassaemia is common in the genotype of αα CS/αα\βN/βN, and clinical manifestations are asymptomatic gene carriers. The genotype of local HbH patients is mainly -α 3.7/-- SEA\βN/βN, and the hematology mainly shows moderate anemia.

8.
Article in Chinese | WPRIM | ID: wpr-864005

ABSTRACT

Refractory pulmonary disease is a respiratory disease that seriously threatens the health of patients, and can lead to impaired pulmonary function and even respiratory failure.Currently, conventional drug therapy does not work well, so it is particularly important to find more effective treatments.Mesenchymal stem cells(MSCs) are a kind of multifunctional stem cells, which can promote the repair of damaged cells by regulating inflammatory and immune reactions and boost the regeneration of damaged tissues through paracrine.Therefore, MSCs are expected to provide a possibility for the treatment of refractory lung pulmonary diseases.

9.
Article in Chinese | WPRIM | ID: wpr-828854

ABSTRACT

OBJECTIVE@#To investigate the effect of β-arrestin1 overexpression on tumor progression in a NCG mouse model bearing T-cell acute lymphocytic leukemia (T-ALL) Molt-4 cell xenograft.@*METHODS@#Molt-4 cells were tagged with firefly-luciferase (F-Luc) by lentiviral infection, and fluorescence intensity of the cells was detected using a luminescence detector. Molt-4 cell lines with β-arrestin1 overexpression or knockdown were constructed by lentivirus infection and injected the tail vein in sub-lethal irradiated NCG mice. Body weight changes and survival time of the xenografted mice were observed, and the progression of T-ALL in the mice was evaluated using an fluorescence imaging system. Sixteen days after xenografting, the mice were euthanatized and tumor cell infiltration was observed in the slices of the liver and spleen.@*RESULTS@#We successfully tagged Molt-4 cells with F-Luc and overexpressed or knocked down β-arrestin1 in the tagged cells. Bioluminescent imaging showed obvious luminescence catalyzed by F-Luc in Molt-4 cells. After injection of Molt-4-Luc cells into irradiated NCG mice, a gradual enhancement of luminescence in the xenografted mice was observed over time, while the body weight of the mice decreased. Compared with the control mice, the mice xenografted with β-arrestin1-overexpressing Molt-4 cells had significantly prolonged survival time ( < 0.001), while the survival time of the mice xenografted with Molt-4 cells with β- arrestin1 knockdown was significantly shortened ( < 0.001). Histological examination revealed fewer infiltrating tumor cells in the liver and spleen of the mice xenografted with β-arrestin1-overexpressing Molt-4 cells in comparison with the mice bearing parental Molt-4 cell xenografts.@*CONCLUSIONS@#β-arrestin1 overexpression suppresses tumor progression in mice bearing Molt-4 cell xenograft.


Subject(s)
Animals , Disease Progression , Heterografts , Humans , Mice , T-Lymphocytes , Transplantation, Heterologous , beta-Arrestin 1
10.
Article in English | WPRIM | ID: wpr-772253

ABSTRACT

Cas1-and-Cas2-mediated new spacer acquisition is an essential process for bacterial adaptive immunity. The process is critical for the ecology of the oral microflora and oral health. Although molecular mechanisms for spacer acquisition are known, it has never been established if this process is associated with the morphological changes of bacteria. In this study, we demonstrated a novel Cas2-induced filamentation phenotype in E. coli that was regulated by co-expression of the Cas1 protein. A 30 amino acid motif at the carboxyl terminus of Cas2 is necessary for this function. By imaging analysis, we provided evidence to argue that Cas-induced filamentation is a step coupled with new spacer acquisition during which filaments are characterised by polyploidy with asymmetric cell division. This work may open new opportunities to investigate the adaptive immune response and microbial balance for oral health.

11.
Article in Chinese | WPRIM | ID: wpr-772053

ABSTRACT

OBJECTIVE@#To establish a stable HEK293T cell line with c.392G>T (p.131G>V) mutation site knockout in gene using CRISPR/Cas9 technique.@*METHODS@#We designed 4 pairs of small guide RNA (sgRNA) for c.392G>T(p.131G>V) mutation site, and constructed exogenous PX458 plasmids expressing Cas9-sgRNA. The plasmids were transfected into HEK293T cells, and the cells expressing GFP fluorescent protein were separated by flow cytometry for further culture. After verification of the knockout efficiency using T7 endonuclease Ⅰ, the monoclonal cells were screened by limiting dilution and DNA sequencing to confirm the knockout. We detected the expressions of mRNA and protein and examined functional changes of the genetically modified cells.@*RESULTS@#We successfully constructed the Cas9-sgRNA exogenous PX458 plasmid based on the c.392G>T(p.131G>V) mutation site of gene. The editing efficiency of the 4 pairs of sgRNA, as detected by T7E1 enzyme digestion, was 6.74%, 12.36%, 12.54% and 2.94%. Sanger sequencing confirmed that the HEK293T cell line with stable knockout of c.392G>T(p.131G>V) was successfully constructed. The genetically modified cells expressed lower levels of mRNA and protein and showed reduced enzyme activity and proliferative capacity and increased apoptosis in response to vitamin K3 treatment.@*CONCLUSIONS@#We successfully constructed a stable HEK293T cell model with gene c.392G>T(p.131G>V) mutation site knockout to facilitate future study of gene repair.


Subject(s)
CRISPR-Cas Systems , HEK293 Cells , Humans , Mutation , Plasmids , RNA, Guide
12.
Article in Chinese | WPRIM | ID: wpr-771886

ABSTRACT

OBJECTIVE@#To explore the correlation of EB virus infection with the prognosis of B-ALL children.@*METHODS@#The peripheral blood of children with newly diagnosed B-ALL admitted in Children's Hospital of Chongqing Medical University from January 2012 to December 2017 were collected, and the EBV DNA in plasma was detected by real-time quantitative PCR. The clinical data of B-ALL children were collected and the correlation of EBV infection with the prognosis of B-ALL children was analyzed.@*RESULTS@#Among 162 B-ALL children, the EBV infection rate was 41.36%. Univariate analysis showed that the B-ALL children with EBV infection had the poor prognosis and higher risk of shorter survival time, as compared with B-ALL children without EBV infection (HR=2.373, 95% CI: 1.129-4.987) (P<0.05), the multivariate analysis showed that the result was consistent with result of univariate analysis indicating that EBV infection was an independent predictor for poor prognosis of B-ALL children.@*CONCLUSION@#The EBV infection may play an important role in the occurrence and progression of B-ALL and is an independent predictor for poor prognosis, therefore the detection of EBV DNA in plasma of B-ALL children possesses an important significance for evaluation of B-ALL children's prognosis.


Subject(s)
B-Lymphocytes , Child , DNA, Viral , Epstein-Barr Virus Infections , Herpesvirus 4, Human , Humans , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Prognosis
13.
Article in Chinese | WPRIM | ID: wpr-771885

ABSTRACT

OBJECTIVE@#To investigate the effect of β-arrestin1 gene on senescence of T-ALL cells and its possible mechanism.@*METHODS@#The bone marrow specimens of T-ALL patients and controls were collected, the expression of β-arrestin1 and β-arrestin1 in the T-ALL patients was detected by RT-PCR and Western blot, respectively, and the relation of β-arrestin1 expression with the clinical pathologic characteristics and the prognosis of T-ALL patients was analyzed statistically. The stable Jurkat cell line with knocked down or overexpressed β-arrestin1 was constructed, the CCK method was used to detect the Jurkat cell number, the β-gal staining was used to analyze the effect of β-arrestin1 on senescence of Jurkat cells, the cross analysis of RNA-Seg data and KEGG data was performed for screening the possible signaling pathway, and Western blot was performed for varifying the key sites of signaling pathway.@*RESULTS@#The β-arrestin1 expression in specimens of T-ALL patients decreased (P<0.01), moreover the β-arrestin1 expression negatively related with peripheral blood cell number (r=-0.601), the blasts in peripheral blood (r=-0.516) and extramedullary infiltration (r=-0.359), while positively related with the response to chemotherapy (r=0.393). The detection of stable Jurkat cell line with knocked-down and overexpressed β-arrestin1 found that the β-arrestin 1 could decrease the Jurkat cell number and accelarate the senescence of Jurkat cells (P<0.05). The cross analysis of RNA-Seg data and KEGG data showed that the senescence of T-ALL cells may be regulated via RAS-P16-PRb-E2F1 by β-arrestin 1. Western bolt confirmed that β-arrestin1 promoted the expression of Ras and p16, and decreased the expression of pRB and E2F1 (P<0.05).@*CONCLUSIONS@#β-arrestin1 accelerates the senescence of Jurkat cells via Ras-p16-pRb-E2F1, and delays the progression in T-ALL, which may provide a new hypothesis for the pathogenesis of T-ALL.


Subject(s)
Cellular Senescence , Humans , Jurkat Cells , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Prognosis , beta-Arrestin 1 , Genetics
14.
Article in Chinese | WPRIM | ID: wpr-702328

ABSTRACT

Objective To systematically review the effects of intracoronary microcatheter agents in the treatment of patients with no-refl ow phenomenon. Methods Databases including Medline, EMbase, the Cochrane Library, CBM, CNKI, VIP and WanFang Data were searched electronically f rom inception to April 2017 for randomized controlled trials (RCTs) about intracoronary agents for no-reflow phenomenon. Two reviewers independently screened literatures, extracted data and assessed the risk of bias of the studies included. Meta-analysis was then performed using RevMan 5.3 software. Results A total of 8 RCTs involving 424 patients were included. The results of Meta-analysis showed that the microcatheter group had significantly better TIMI flow grade[RR=0.38,95%CI(0.27,0.52), P<0.000 01],TIMI myocardial perf usion grade[RR=0.35,95%CI(0.23,0.55),P<0.000 01],corrected TIMI f rame count[MD=-9.99,95%CI(-13.22,-6.76)P<0.000 01]and hypotension[RR=0.57,95%CI(0.35, 0.90),P=0.02] than those of the guiding catheter group. There was no statistical difference between the two groups in short period major adverse cardiovascular events and left ventricular ejection fraction.Conclusions Current evidence shows that intracoronary microcatheter agents could improve blood flow in patients with no-reflow phenomenon and has good safety. Due to the limited quantity and quality of the included studies, the above conclusion still needs to be verified by carrying out more high-quality RCTs.

15.
Article in Chinese | WPRIM | ID: wpr-692685

ABSTRACT

Objective To explore the diagnostic value of serum lipoprotein(a)[Lp(a)]in cerebral infarction by a model of Logistic regression and receiver operating characteristic(ROC)curve.Methods A total of 316 patients with cerebral thrombosis from Foshan Hospital Affiliated to Southern Medical University were col-lected.According to the diagnostic criteria,the patients were divided into cerebral thrombosis group(196 ca-ses)and non-cerebral thrombosis group(120 cases).All the subjects were tested for Lp(a)by immune turbi-dimetry.To evaluate the diagnostic value of Lp(a)by applying logistic regression model,drawing ROC curves and calculating the area under the curve(AUC).Results The P25,P50,P75of Lp(a)in cerebral thrombosis group and non-cerebral thrombosis group were 97.23,238.22,430.01 and 29.80,92.27,233.86,the average rank were 185.42 and 114.52,the differences in the two groups were significant(P<0.05).Logistic regres-sion showed that the correlation between Lp(a)level and cerebral thrombosis was positive,the partial regres-sion coefficient(B)was 0.005,Wald value was 31.295.It suggested that when the levels of Lp(a)was higher the risk of cerebral thrombosis increased.The most valuable diagnosis level was 305.80 mg/L.And the area under the ROC curve(AUC)was 0.724,which has moderate diagnostic efficacy.Diagnostic specificity was 91.7%,misdiagnosis rate was 8.3%,negative predictive value was 48.7%,sensitivity was 40.8%,omission rate was 59.2%,positive predictive value was 88.9%.Conclusion The level of serum Lp(a)has high diag-nostic specificity for the diagnosis of cerebral thrombosis.

16.
Article in Chinese | WPRIM | ID: wpr-692626

ABSTRACT

Objective To analyze the comparability of immune qualitative items results in the clinical labo-ratory department of partial tertiary hospital and second-class hospitals of Chongqing City during 2013-2016 to lay the foundation for realizing the results mutual recognition of immune qualitative items .Methods The basic situation survey and inquiry were conducted by the questionnaire survey and spot-supervision modes .The comparison and analysis of fresh blood samples for hepatitis Bimmune markers and autoantibody items were continuously performed from 2013 to 2016 .The problems were found by the investigation guidance and fresh blood samples comparison by stages ,and then continuously improving the test quality was performed .Results The investigation and analysis found that the development in immune sub-professional group of hospital clinical laboratory department in Chongqing City was imbalance ,the resource allocation had large difference and the coverage rate of internal quality items urgently needed to be increased .In the comparison of fresh blood samples during 2013-2016 ,the items of HBsAg and HBsAb had good consistency ,followed by HBeAg . The average scores of item passing rate during 2013-2016 were 99 .23% ,100 .00% ,96 .40% and 98 .72% for HBsAg ;100 .00% ,99 .30% ,97 .00% and 98 .46% for anti-HBs ;98 .50% ,92 .40% ,92 .70% and 97 .69% for HBeAg .The results consistency of HBeAb and HBcAb was poorer than that of the former 3 items due to large difference in the reagent ,instrument and methodology .The consistency rate in the comparison the karyotype , negative and positive in the autoantibody items was 100 .00% ,but there was few laboratories carrying out this items(<15) .Conclusion The quality of immune mutual recognition items of tertiary hospital laboratories is good ,and the test quality of partial items in the second-class hospital laboratory still needs to be further in-creased;through the immune professional investigation and analysis ,and the comparison of fresh blood sam-ples ,standardizing the internal quality control and external quality assessment can increase the comparability and accuracy of inter-hospital test results mutual recognition to lay the foundation for promoting the hierarchi-cal diagnosis and treatment .

17.
Article in Chinese | WPRIM | ID: wpr-690478

ABSTRACT

<p><b>OBJECTIVE</b>To explore the key genes in T-cell acute lymphoblastic leukemia (T-ALL) using bioinformatics method to better understand the pathogenic mechanisms of T-ALL.</p><p><b>METHODS</b>The gene expression profiles of GSE14317 were obtained from Gene Expression Omnibus database. The differentially expressed genes (DEGs) in T-ALL were analyzed using R package Limma. The online analysis tool DAVID was used to perform the functional and pathway enrichment analysis. The protein-protein interaction network was constructed by STRING and visualized by Cytoscape. Based on the JASPAR database, the transcription factors (TFs) of the hub genes were obtained. RT-PCR was used to test the mRNA expression level of the key genes.</p><p><b>RESULTS</b>A total of 1443 DEGs were identified, including 800 up-regulated genes and 643 down-regulated genes. These DEGs were significantly enriched in the cell cycle, hematopoietic cell lineage, cytokine-cytokine receptor interaction and T cell receptor signaling pathway. The top 10 hub genes identified from the PPI networks included CDK1, PIK3R1, CCNB1, CCNA2, CDC20, JUN, GNG11, PLK1, PCNA and CCNB2, which were enriched in chemokine signaling pathway, ubiquition mediated proteolysis and cell cycle. In the TF-target gene network, 42 differentially expressed TFs were identified, among which ELF5, HIC2 and MEISI had binding sites with 9 of the candidate hub genes. RT-PCR showed that the mRNA expression level of all the candidate hub genes except for GNG11 were consistent with the gene expression profiles.</p><p><b>CONCLUSION</b>The hub genes CDK1, PIK3R1, CCNB1, CCNA2, CDC20, JUN, PLK1, PCNA, CCNB2, ELF5, HIC2 and MEISI participate in the occurrence of T-ALL. Our finding provides new insights into the pathogenesis of T-ALL.</p>

18.
Article in Chinese | WPRIM | ID: wpr-737216

ABSTRACT

Ezetimibe was reported to pharmacologically defend against oxidative stress.This study was designed to investigate whether ezetimibe can protect against the oxidative stress induced by oxidized low-density lipoprotein (oxLDL) in vitro and the underlying mechanism.Human umbilical vein endothelial cells (HUVECs) were pretreated with ezetimibe and then exposed to oxLDL for 24 h.TUNEL assay and detection for the protein levels of cleaved caspase-3,Bcl-xl and Bcl-2 were employed to assess the oxLDL-induced endothelial apoptosis.Intracellular reactive oxygen species (ROS) generation was evaluated by measuring dichlorofluorescein (DCF) fluorescence.The activities of endothelial antioxidant enzymes [superoxide dismutase (SOD) and catalase] were tested via an enzymatic assay.The mitochondrial membrane potential (MMP) was monitored by flow cytometry using JC-1 staining.Phosphorylation levels of glycogen synthase kinase-3β (p-GSK-3β) and Akt (p-Akt),as well as total GSK-3β and Akt were determined by Western blotting.The results showed that ezetimibe treatment inhibited HUVECs apoptosis,intracellular ROS production,and enhanced antioxidant enzyme activities elicited by oxLDL.HUVECs exposed to oxLDL alone had reduced mitochondrial function,while ezetimibe pre-intervention could significantly rescue the MMP.Furthermore,the protein levels of p-GSK-3β and p-Akt in ezetimibe-pretreated HUVECs were markedly increased as compared with those in oxLDL-induced HUVECs.However,no significant effect on total GSK-3β and Akt was found in ezetimibe-pretreated HUVECs.Taken together,it was concluded that ezetimibe protects against oxLDL-induced oxidative stress through restoring the MMP,which may be mediated by Akt-dependent GSK-3β phosphorylation.

19.
Article in Chinese | WPRIM | ID: wpr-735748

ABSTRACT

Ezetimibe was reported to pharmacologically defend against oxidative stress.This study was designed to investigate whether ezetimibe can protect against the oxidative stress induced by oxidized low-density lipoprotein (oxLDL) in vitro and the underlying mechanism.Human umbilical vein endothelial cells (HUVECs) were pretreated with ezetimibe and then exposed to oxLDL for 24 h.TUNEL assay and detection for the protein levels of cleaved caspase-3,Bcl-xl and Bcl-2 were employed to assess the oxLDL-induced endothelial apoptosis.Intracellular reactive oxygen species (ROS) generation was evaluated by measuring dichlorofluorescein (DCF) fluorescence.The activities of endothelial antioxidant enzymes [superoxide dismutase (SOD) and catalase] were tested via an enzymatic assay.The mitochondrial membrane potential (MMP) was monitored by flow cytometry using JC-1 staining.Phosphorylation levels of glycogen synthase kinase-3β (p-GSK-3β) and Akt (p-Akt),as well as total GSK-3β and Akt were determined by Western blotting.The results showed that ezetimibe treatment inhibited HUVECs apoptosis,intracellular ROS production,and enhanced antioxidant enzyme activities elicited by oxLDL.HUVECs exposed to oxLDL alone had reduced mitochondrial function,while ezetimibe pre-intervention could significantly rescue the MMP.Furthermore,the protein levels of p-GSK-3β and p-Akt in ezetimibe-pretreated HUVECs were markedly increased as compared with those in oxLDL-induced HUVECs.However,no significant effect on total GSK-3β and Akt was found in ezetimibe-pretreated HUVECs.Taken together,it was concluded that ezetimibe protects against oxLDL-induced oxidative stress through restoring the MMP,which may be mediated by Akt-dependent GSK-3β phosphorylation.

20.
Basic & Clinical Medicine ; (12): 939-944, 2017.
Article in Chinese | WPRIM | ID: wpr-612014

ABSTRACT

Objective To establish a method for detection of serum urocanic acid (UCA) by high performance liquid chromatography (HPLC),and explore the clinical significance of serum UCA concentration for children acute leukemia.Methods The chromatographic conditions of HPLC were set up and optimized,and the linearity of standard curve,precision,accuracy and stability were validated.Then the serum from ninety acute leukemia children and ninety non-tumor blood disease children was collected,the concentration of serum UCA was detected with HPLC,and the differences of two groups were compared to study the clinical significance of UCA in children acute leukemia.Results The HPLC method for detecting serum UCA was successfully established and optimized.The standard curves of trans-UCA and cis-UCA both showed good linearities(R2=0.999 6 and 0.999 9) at the condition of the mobile phase of acetonitrile-20 mmol/L KH2PO4,pH 3.7(5:95,V/V),flow rate of 1.2 mL/min,detection wavelength of 264 nm in HPLC.The relative standard deviation RSD% of intra-assay and inter-assay were lower than 5%.Compared with non-tumor blood disease,the serum concentration of cis urocanic acid (cis-UCA) and trans urocanic acid (trans-UCA) of children with acute leukemia were significantly increased (P<0.001).Compared with cis-UCA,trans-UCA was more valuable for risk classification of acute lymphoblastic leukemia (ALL).Conclusions HPLC is a good technology to titrate of UCA in serum.The concentration of serum UCA in children with acute leukemia may provide the clues for diagnosis and prognosis,with important clinical significance.

SELECTION OF CITATIONS
SEARCH DETAIL