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1.
Article in Chinese | WPRIM | ID: wpr-879584

ABSTRACT

OBJECTIVE@#To detect fusion gene with pathological significance in a patient with refractory and relapsed acute B cell lymphoblastic leukemia (B-ALL) and to explore its laboratory and clinical characteristics.@*METHODS@#Transcriptome sequencing was used to detect potential fusion transcripts. Other laboratory results and clinical data of the patient were also analyzed.@*RESULTS@#The patient was found to harbor TCF3 exon 17-ZNF384 exon 7 in-frame fusion transcript. The minimal residual disease (MRD) has remained positive after multiple chemotherapy protocols including CD19-, CD22- targeted chimeric antigen receptor T cells immunotherapy. The patient eventually achieved complete remission and sustained MRD negativity after allogeneic hemopoietic stem cell transplantation (allo-HSCT).@*CONCLUSION@#Transcriptome sequencing can effectively detect potential fusion genes with clinical significance in leukemia. TCF3-ZNF384 positive B-ALL has unique laboratory and clinical characteristics, may not well respond to chemotherapy and immunotherapy, and is more likely to relapse. Timely allo-HSCT treatment may help such patients to achieve long-term disease-free survival. TCF3-ZNF384 positive B-ALL is not uncommon in pediatric patients but has not been effectively identified.


Subject(s)
B-Lymphocytes , Basic Helix-Loop-Helix Transcription Factors/genetics , Child , Hematopoietic Stem Cell Transplantation , Humans , Laboratories , Precursor Cell Lymphoblastic Leukemia-Lymphoma/therapy , Trans-Activators/genetics , Transcriptome
2.
Arq. neuropsiquiatr ; 77(3): 166-173, Mar. 2019. tab, graf
Article in English | LILACS | ID: biblio-1001345

ABSTRACT

ABSTRACT It is currently unknown how genetic factors may influence the clinical course of multiple sclerosis (MS). Objective: We examined the impact of CIITA polymorphisms −168A/G (rs3087456) and +1614G/C (rs4774) on the risk of disability progression, severity and on responses to first-line immunomodulator treatments. Methods: Genomic DNA was extracted from blood samples. We used ABI3730xl and GeneMapper v.4.0 software to identify genotype variations. All patients were followed up and clinically reassessed at three-month intervals. Disability progression was measured by the Expanded Disability Status Scale and disease severity by the Multiple Sclerosis Spasticity Scale (MSSS). Results: We included 37 men and 80 women. We found no evidence regarding the influence of the single nucleotide polymorphisms studied in the Expanded Disability Status Scale or therapeutic response of the evaluated drugs. We performed a logistic regression analysis with the MSSS and found that a less severe MS course was associated with wild type CIITA −168AA and CIITA +1614GG, as the chance of the patient progressing to MSSS2 and MSSS3 decreased in 61% and 75% with CIITA −168AA and 66% and 75% with CIITA +1614GG, respectively (p < 0.0001). Although less significant, the CIITA +1614 GC also pointed to a less severe MS course and the chance of the patient progressing to MSSS3 decreased 79% (p = 0.015). We also observed that the CIITA −168GG genotype was more frequent in MSSS2 and MSSS3 and had 40% lower odds ratio to becoming more severe MS. Conclusion: These data suggest that CIITA −168AA, CIITA +1614GG and CIITA +1614 GC polymorphisms may be associated with a better MS clinical course. This knowledge may be useful for a better understanding of MS and its therapeutic management.


RESUMO Atualmente não se sabe como os fatores genéticos podem influenciar o curso clínico da esclerose múltipla (EM). Objetivo: Examinamos o impacto dos polimorfismos CIITA −168A/G (rs3087456) e CIITA +1614G/C (rs4774) no risco de progressão da incapacidade, gravidade e resposta aos tratamentos imunomoduladores de primeira linha. Métodos: O DNA genômico foi extraído de amostras de sangue. Utilizamos o software ABI3730xl e GeneMapper v.4.0 (Applied Biosystems) para identificar variações genotípicas. Todos os pacientes foram acompanhados e reavaliados clinicamente em intervalos de três meses. A progressão da incapacidade foi medida pela EDSS e a gravidade da doença pelo MSSS. Resultados: Incluímos 37 homens e 80 mulheres. Não encontramos evidências sobre a influência dos SNPs estudados no EDSS e na resposta terapêutica aos fármacos avaliados. Realizamos uma análise de regressão logística com o MSSS e observamos uma evolução menos grave da EM associada aos tipos selvagens CIITA −168AA e CIITA +1614GG, pois a chance do paciente atingir MSSS2 e MSSS3 diminuiu em 61%/75%, e 66/75% respectivamente (p < 0,0001). Embora menos significativo, o CIITA +1614GC também foi relacionado com evolução menos grave da EM e a chance do paciente atingir o MSSS3 diminuiu 79% (p = 0,015). Nós também observamos que o genótipo CIITA −168GG foi mais frequente no MSSS2 e MSSS3 e teve uma razão de chance 40% menor para atingir forma mais grave da EM. Conclusão: Estes dados sugerem que os polimorfismos CIITA −168AA, CIITA +1614GG e CIITA +1614GC podem estar associados a um melhor curso clínico da EM. Este conhecimento pode ser útil para uma melhor compreensão da EM e o seu manejo terapêutico.


Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Young Adult , Nuclear Proteins/genetics , Trans-Activators/genetics , Disease Progression , Polymorphism, Single Nucleotide/genetics , Multiple Sclerosis/genetics , Time Factors , Severity of Illness Index , Logistic Models , Retrospective Studies , Interferon-beta/therapeutic use , Disability Evaluation , Kaplan-Meier Estimate , Genetic Association Studies , Glatiramer Acetate/therapeutic use , Gene Frequency , Genotype , Immunologic Factors/therapeutic use , Multiple Sclerosis/mortality , Multiple Sclerosis/drug therapy
3.
Article in English | WPRIM | ID: wpr-771436

ABSTRACT

OBJECTIVE@#To investigate the effect of a modified Wuzi Yanzong Pill (, WZYZP) on the male rats' testis after microwave radiation, as well as its potential mechanism.@*METHODS@#Forty-five male rats were randomly assigned to three groups: the control group, the radiation group, and the WZYZP group. The rats in the radiation group and WZYZP group were exposed to microwave radiation for 15 min once, while the rats in the control group were not exposed to any radiation. The rats in the WZYZP group were given a modified of WZYZP by gavage daily for 7 days. Apoptosis in the testis was evaluated using terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay. Histopathological alterations of the testis were observed by haematoxylin-eosin (HE) staining. Tat-interactive protein, 60kD (Tip60) and p53 expressions were determined by Western blotting.@*RESULTS@#The apoptosis index (AI) in the radiation group was higher than that of the WZYZP group and control group on day 1 (D1), day 7 (D7) day 14 (D14) after radiation (P<0.05). The seminiferous tubules were of normal morphology in the control group. In the radiation group, the partial seminiferous tubules were collapsed, basement membranes of the seminiferous epithelia became detached. WZYZP could restore the morphological changes. There was no expression of Tip60 among the three groups on D7 and D14. The expression of p53 was higher in the radiation group than in the control group (P<0.05). WZYZP could down-regulate the rising p53 induced by radiation on D7 and D14 (P<0.05).@*CONCLUSION@#A modified WZYZP may affect germ cells, and its protective effects may partly result from its ability to intervene in Tip60 mediated apoptosis.


Subject(s)
Animals , Apoptosis , Drugs, Chinese Herbal , Pharmacology , Male , Microwaves , Rats, Wistar , Testis , Metabolism , Pathology , Radiation Effects , Trans-Activators , Metabolism , Tumor Suppressor Protein p53 , Metabolism
4.
Article in Chinese | WPRIM | ID: wpr-777523

ABSTRACT

A stable hepatoma cell line(Hep G2 cell) insulin resistance model was established and used to analyze the effect of effective components of Mori Folium in alleviating insulin resistance,and preliminary explore the mechanism for alleviating insulin resistance. The Hep G2 insulin action concentration and the duration of action were investigated using the glucose oxidase method(GOD-POD method) to establish a stable Hep G2 insulin resistance model. Normal control group,model group,Mori Folium polysaccharide group,Mori Folium flavonoid group and rosiglitazone group were divided to determine the glucose consumption. The effect of Mori Folium effective components on Hep G2 insulin resistance was analyzed. The mRNA expressions of JNK,IRS-1 and PDX-1 in each group were detected by Real-time quantitative PCR(qRT-PCR). The protein expressions of p-JNK,IRS-1 and PDX-1 were detected by Western blot. And the mechanism of effective components of Mori Folium in alleviating insulin resistance was investigated. The results showed that the glucose consumption was significantly decreased in the insulin resistance cells after incubation with 25. 0 mg·L-1 insulin for 36 h(P<0. 01),and the model was relatively stable within 36 h. Mori Folium polysaccharides and flavonoids all alleviated insulin resistance,among which Mori Folium flavonoids had better effect in alleviating Hep G2 insulin resistance(P<0. 05). The qRT-PCR analysis showed that Mori Folium polysaccharides and flavonoids could inhibit JNK and IRS-1 mRNA expressions,while enhancing PDX-1 mRNA expression. Western blot analysis displayed that Mori Folium polysaccharides and flavonoids could inhibit p-JNK and IRS-1 protein expressions,while enhancing PDX-1 protein expression. Mori Folium polysaccharides and flavonoids can alleviate insulin resistance in Hep G2 cells,and its mechanism may be the alleviation of insulin resistance by inhibiting JNK signaling pathway.


Subject(s)
Drugs, Chinese Herbal , Pharmacology , Glucose , Hep G2 Cells , Homeodomain Proteins , Metabolism , Humans , Insulin , Insulin Receptor Substrate Proteins , Metabolism , Insulin Resistance , MAP Kinase Kinase 4 , Metabolism , MAP Kinase Signaling System , Morus , Chemistry , Plant Leaves , Chemistry , Trans-Activators , Metabolism
5.
Article in Chinese | WPRIM | ID: wpr-775079

ABSTRACT

OBJECTIVE@#To investigate the effect and molecular mechanism of interferon-α (INF-α) on the apoptosis of the mouse podocyte cell line MPC5 induced by hepatitis B virus X (HBx) protein.@*METHODS@#MPC5 cells were transfected with the pEX plasmid carrying the HBx gene. RT-PCR was used to measure the mRNA expression of HBx at different time points. MPC5 cells were divided into 4 groups: control group (MPC5 cells cultured under normal conditions), INF-α group (MPC5 cells cultured with INF-α), HBx group (MPC5 cells induced by HBx), and HBx+INF-α group (MPC5 cells induced by HBx and cultured with INF-α). After 48 hours of intervention under different experimental conditions, flow cytometry was used to measure the apoptosis of MPC5 cells, and quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of slit diaphragm-related proteins (nephrin, CD2AP, and synaptopodin) and the cytoskeleton-related protein transient receptor potential cation channel 6 (TRPC6).@*RESULTS@#MPC5 cells transfected by pEX-HBx had the highest expression of HBx mRNA at 48 hours after transfection (P<0.05). Compared with the control, INF-α and HBx+INF-α groups, the HBx group had a significant increase in the apoptosis rate of MPC5 cells (P<0.05). Compared with the control and INF-α groups, the HBx group had significant reductions in the mRNA and protein expression of nephrin, synaptopodin, and CD2AP and significant increases in the mRNA and protein expression of TRPC6 (P<0.05). Compared with the HBx group, the HBx+INF-α group had significant increases in the mRNA and protein expression of nephrin, synaptopodin, and CD2AP and significant reductions in the mRNA and protein expression of TRPC6 (P<0.05).@*CONCLUSIONS@#INF-α can inhibit the apoptosis of podocytes induced by HBx, possibly through improving the abnormal expression of slit diaphragm-related proteins (CD2AP, nephrin, and synaptopodin) and cytoskeleton-related protein (TRPC6) induced by HBx.


Subject(s)
Animals , Apoptosis , Hepatitis B virus , Interferon-alpha , Mice , Podocytes , Trans-Activators
6.
Article in Chinese | WPRIM | ID: wpr-813008

ABSTRACT

To examine the expression of vasohibin-1, metastasis-associated in colon cancer-1 (MACC1) and KAI1 proteins in serous ovarian cancer and their clinical significance.
 Methods: In 124 specimens of serous ovarian cancer (serous ovarian cancer group) and 30 specimens of ovarian serous cystadenoma (ovarian serous cystadenoma group), the expression of vasohibin-1, MACC1 and KAI1 protiens were detected by immunohistochemistry ElivisionTM method.
 Results: In the serous ovarian cancer group, the positive rates of vasohibin-1 and MACC1 proteins were 48.4% and 58.1%, respectively, which were both higher than those in the ovarian serous cystadenoma group (10.0% and 13.3%, respectively); while the positive rate of KAI1 protein in the serous ovarian cancer group was 33.9%, which was lower than that in the ovarian serous cystadenoma group (86.7%), there were significant differences between the 2 groups (all P<0.05). In the serous ovarian cancer group, the expression of the 3 proteins were closely related to the pathological grade, Federation International of Gynecology and Obstetrics (FIGO) stage and pelvic lymph node metastasis (all P<0.05). The KAI1 protein was negatively correlated with the levels of vasohibin-1 and MACC1 (r=-0.500, -0.600, respectively, both P<0.01); while there was a positive correlation between the vasohibin-1 and the MACC1 (r=0.518, P<0.01). Kaplan-Meier survival analysis showed that the over-expression of vasohibin-1, MACC1 and the low-expression of KAI1 protein were related to the survival rates (all P<0.05). Multi-factor analysis showed that the expression of vasohibin-1, KAI1 protein and the FIGO stage were independent prognosis factors for radical operation of serous ovarian cancer (RR=2.185, 3.893, 0.413; 95% CI=1.263-3.779, 2.190-6.921, 0.251-0.681; all P<0.05).
 Conclusion: The up-regulation of vasohibin-1, MACC1 and down-regulation of KAI1 in serous ovarian cancer are related to the tumor differentiation, clinical stage, metastasis and prognosis. Combined detection of these indexes is useful in predicting the progression and prognosis of serous ovarian cancer.


Subject(s)
Carcinoma, Ovarian Epithelial , Cell Cycle Proteins , Colonic Neoplasms , Female , Humans , Kangai-1 Protein , Neoplasm Staging , Ovarian Neoplasms , Prognosis , Trans-Activators , Transcription Factors
7.
Article in Chinese | WPRIM | ID: wpr-775660

ABSTRACT

BACKGROUND@#Non-small cell lung cancer (NSCLC) is a kind of lung cancer, because its high incidence has been concerned. Therefore, it has great significance to reveal the pathogenesis of NSCLC. As a transcriptional regulatory factor, MATF-A plays an important role in the development of multiple tumors, can regulate the migration process of a variety of tumor cells. HOTAIR is a long non-coding RNA (LncRNA) found in recent years, which expresses abnormally in multiple tumors and is involved in the proliferation and migration of multiple tumors. The aim of this study is to explore the role of MRTF-A through HOTAIR to regulate the proliferation and migration of NSCLC cell A549 cell.@*METHODS@#We constructed the overexpression plasmid and interfering plasmid of MRTF-A, and detected the effect of MRTF-A on the proliferation and migration of A549 cells by CCK8 and wound healing methods respectively. Then, we designed the siRNA of HOTAIR to detect its effect on the proliferation and migration of A549 cells. Through qRT-PCR, we detected the effect of MRTF-A on HOTAIR expression. Finally, we constructed HOTAIR's promoter, and detect the effect of MRTF-A on HOTAIR promoter activity by luciferase reporter gene test.@*RESULTS@#Overexpression of MRTF-A promotes the proliferation and migration of A549 cells, while silent MRTF-A inhibits its proliferation and migration. Next, we found that interfered HOTAIR expression inhibited the proliferation of A549 cells. We found that MRTF-A could influence the expression of HOTAIR and regulate the activity of HOTAIR promoter.@*CONCLUSIONS@#MRTF-A regulates the proliferation and migration of A549 cell through HOTAIR.


Subject(s)
A549 Cells , Carcinoma, Non-Small-Cell Lung , Genetics , Metabolism , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Humans , Promoter Regions, Genetic , RNA, Long Noncoding , Genetics , Metabolism , Trans-Activators , Genetics , Metabolism
8.
Braz. j. infect. dis ; 22(2): 129-136, Mar.-Apr. 2018. tab, graf
Article in English | LILACS | ID: biblio-951633

ABSTRACT

ABSTRACT Introduction: Biofilm production is an important mechanism for the survival of Pseudomonas aeruginosa and its relationship with antimicrobial resistance represents a challenge for patient therapeutics. P. aeruginosa is an opportunistic pathogen frequently associated to nosocomial infections, especially in imunocompromised hosts. Objectives: Analyze the phenotypic biofilm production in P. aeruginosa isolates, describe clonal profiles, and analyze quorum sensing (QS) genes and the occurrence of mutations in the LasR protein of non-biofilm producing isolates. Methods: Isolates were tested for biofilm production by measuring cells adherence to the microtiter plates. Clonal profile analysis was carried out through ERIC-PCR, QS genes were by specific PCR. Results: The results showed that 77.5% of the isolates were considered biofilm producers. The results of genotyping showed 38 distinct genetic profiles. As for the occurrence of the genes, 100% of the isolates presented the lasR, rhlI and rhlR genes, and 97.5%, presented the lasI gene. In this study nine isolates were not biofilm producers. However, all presented the QS genes. Amplicons related to genes were sequenced in three of the nine non-biofilm-producing isolates (all presenting different genetic similarity profile) and aligned to the sequences of those genes in P. aeruginosa strain PAO1 (standard biofilm-producing strain). Alignment analysis showed an insertion of three nucleotides (T, C and G) causing the addition of an amino acid valine in the sequence of the LasR protein, in position 53. Conclusion: The modeling of the resulting LasR protein showed a conformational change in its structure, suggesting that this might be the reason why these isolates are unable to produce biofilm.


Subject(s)
Humans , Pseudomonas aeruginosa/physiology , Pseudomonas Infections/microbiology , Bacterial Proteins/genetics , Trans-Activators/genetics , Biofilms/growth & development , Biofilms/drug effects , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/chemistry , Pseudomonas Infections/drug therapy , Bacterial Proteins/chemistry , Trans-Activators/chemistry , Polymerase Chain Reaction/methods , Cross Infection , Drug Resistance, Multiple, Bacterial , Anti-Infective Agents/pharmacology , Anti-Bacterial Agents/pharmacology
9.
Braz. j. med. biol. res ; 51(9): e7588, 2018. tab, graf
Article in English | LILACS | ID: biblio-951758

ABSTRACT

Previous studies suggested that chromodomain helicase DNA-binding proteins (CHDs), including CHD 1-8, were associated with several human diseases and cancers including lymphoma, liver cancer, colorectal cancer, stomach cancer, etc. To date, little research on CHD 9 in human cancers has been reported. In this study, we assessed the prognostic value of CHD 9 in patients with colorectal cancer (CRC). We screened for CHD 9 expression using immunohistochemical analysis in 87 surgical CRC specimens and found that the expression was upregulated in 81.5% of the cases, while 7.4% were decreased; in the remaining 11.1% of the cases, levels were not altered. Kaplan-Meier analysis showed that patients with high CHD 9 expression had better prognosis than those with low CHD 9 expression (54.5 vs 32.1%, P=0.034). Subsequently, Cox multi-factor survival regression analysis revealed that expression of CHD 9 protein was an independent predictor for CRC, with a hazard ratio of 0.503 (P=0.028). In addition, we found that CHD 9 expression was positively correlated with MSH2 (rs=0.232, P=0.036). We speculated that CHD9 might be a putative tumor suppressor gene, and could inhibit the development of CRC by participating in DNA repair processes. Our findings suggest that CHD 9 could be a novel prognostic biomarker and a therapeutic target for CRC. Further studies are needed to detect the effect of CHD 9 on cellular function and the expression of mismatch repair genes.


Subject(s)
Humans , Male , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Transcription Factors/metabolism , Colorectal Neoplasms/metabolism , Biomarkers, Tumor/metabolism , DNA-Binding Proteins/metabolism , Prognosis , Transcription Factors/genetics , Immunohistochemistry , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Biomarkers, Tumor/genetics , Gene Expression Regulation, Neoplastic , Trans-Activators , DNA Helicases , DNA-Binding Proteins/genetics , Kaplan-Meier Estimate , Neoplasm Staging
10.
Braz. j. microbiol ; 48(1): 118-124, Jan.-Mar. 2017. tab, graf
Article in English | LILACS | ID: biblio-839334

ABSTRACT

Abstract Staphylococcus spp. play an important role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most relevant species due to the production of virulence factors such as slime, which is required for biofilm formation. This study aimed to evaluate biofilm production and its possible relation to beta-lactamic resistance in 20 S. aureus isolates from bovine mastitic milk. The isolates were characterized by pheno-genotypic and MALDI TOF-MS assays and tested for genes such as icaA, icaD, bap, agr RNAIII, agr I, agr II, agr III, and agr IV, which are related to slime production and its regulation. Biofilm production in microplates was evaluated considering the intervals determined along the bacterial growth curve. In addition, to determine the most suitable time interval for biofilm analysis, scanning electron microscopy was performed. Furthermore, genes such as mecA and blaZ that are related to beta-lactamic resistance and oxacillin susceptibility were tested. All the studied isolates were biofilm producers and mostly presented icaA and icaD. The Agr type II genes were significantly prevalent. According to the SEM, gradual changes in the bacterial arrangement were observed during biofilm formation along the growth curve phases, and the peak was reached at the stationary phase. In this study, the penicillin resistance was related to the production of beta-lactamase, and the high minimal bactericidal concentration for cefoxitin was possibly associated with biofilm protection. Therefore, further studies are warranted to better understand biofilm formation, possibly contributing to our knowledge about bacterial resistance in vivo.


Subject(s)
Animals , Female , Staphylococcal Infections/microbiology , Staphylococcus aureus/drug effects , Staphylococcus aureus/physiology , Biofilms , beta-Lactam Resistance , Mastitis, Bovine/microbiology , Anti-Bacterial Agents/pharmacology , Staphylococcus aureus/ultrastructure , Bacterial Proteins/genetics , Cattle , Microbial Sensitivity Tests , Trans-Activators/genetics , Proteome , Virulence Factors/genetics , Proteomics/methods , Genetic Association Studies
11.
Blood Research ; : 106-111, 2017.
Article in English | WPRIM | ID: wpr-62220

ABSTRACT

BACKGROUND: Adult T-cell leukemia/lymphoma (ATLL) is an aggressive malignancy with very poor prognosis and short survival, caused by the human T-lymphotropic virus type-1 (HTLV-1). The HTLV-1 biomarkers trans-activator x (TAX) and HTLV-1 basic leucine zipper factor (HBZ) are main oncogenes and life-threatening elements. This study aimed to assess the role of the TAX and HBZ genes and HTLV-1 proviral load (PVL) in the survival of patients with ATLL. METHODS: Forty-three HTLV-1-infected individuals, including 18 asymptomatic carriers (AC) and 25 ATLL patients (ATLL), were evaluated between 2011 and 2015. The mRNA expression of TAX and HBZ and the HTLV-1 PVL were measured by quantitative PCR. RESULTS: Significant differences in the mean expression levels of TAX and HBZ were observed between the two study groups (ATLL and AC, P=0.014 and P=0.000, respectively). In addition, the ATLL group showed a significantly higher PVL than AC (P=0.000). There was a significant negative relationship between PVL and survival among all study groups (P=0.047). CONCLUSION: The HTLV-1 PVL and expression of TAX and HBZ were higher in the ATLL group than in the AC group. Moreover, a higher PVL was associated with shorter survival time among all ATLL subjects. Therefore, measurement of PVL, TAX, and HBZ may be beneficial for monitoring and predicting HTLV-1-infection outcomes, and PVL may be useful for prognosis assessment of ATLL patients. This research demonstrates the possible correlation between these virological markers and survival in ATLL patients.


Subject(s)
Adult , Biomarkers , Human T-lymphotropic virus 1 , Humans , Leucine Zippers , Leukemia-Lymphoma, Adult T-Cell , Oncogenes , Polymerase Chain Reaction , Prognosis , RNA, Messenger , T-Lymphocytes , Taxes , Trans-Activators
12.
Immune Network ; : 410-423, 2017.
Article in English | WPRIM | ID: wpr-10876

ABSTRACT

Neutrophils and eosinophils, 2 prominent granulocytes, are commonly derived from myelocytic progenitors through successive stages in the bone marrow. Our previous genome-wide transcriptomic data unexpectedly showed that genes encoding a multitude of neutrophil primary granule proteins (NPGPs) were markedly downregulated during the end period of eosinophilic terminal differentiation when cord blood (CB) cluster of differentiation (CD) 34+ cells were induced to differentiate toward the eosinophil lineage during a 24-day culture period. Accordingly, this study aimed to examine whether NPGP genes were expressed on the way to eosinophil terminal differentiation stage and to compare their expression kinetics with that of genes encoding eosinophil-specific granule proteins (ESGPs). Transcripts of all NPGP genes examined, including proteinase 3, myeloperoxidase, cathepsin G (CTSG), and neutrophil elastase, reached a peak at day 12 and sharply declined thereafter, while transcript of ESGP genes including major basic protein 1 (MBP1) attained maximum expression at days 18 or 24. Growth factor independent 1 (GFI1) and CCAAT/enhancer-binding protein α (C/EBPA), transactivators for the NPGP genes, were expressed immediately before the NPGP genes, whereas expression of C/EBPA, GATA1, and GATA2 kinetically paralleled that of eosinophil granule protein genes. The expression kinetics of NPGPs and ESGPs were duplicated upon differentiation of the eosinophilic leukemia cell line (EoL-1) immature eosinophilic cells. Importantly, confocal image analysis showed that CTSG was strongly coexpressed with MBP1 in differentiating CB eosinophils at days 12 and 18 and became barely detectable at day 24 and beyond. Our results suggest for the first time the presence of an immature stage where eosinophils coexpress NPGPs and ESGPs before final maturation.


Subject(s)
Bone Marrow , Cathepsin G , Cell Line , Eosinophils , Fetal Blood , Granulocytes , Hypereosinophilic Syndrome , Kinetics , Leukocyte Elastase , Myeloblastin , Neutrophils , Peroxidase , Trans-Activators
14.
Chinese Medical Journal ; (24): 1355-1362, 2016.
Article in English | WPRIM | ID: wpr-290072

ABSTRACT

<p><b>BACKGROUND</b>The acute myeloid leukemia 1 (AML1)-eight-twenty-one (ETO) fusion protein generated by the t(8;21)(q22;q22) translocation is considered to display a crucial role in leukemogenesis in AML. By focusing on the anti-leukemia effects of eyes absent 4 (EYA4) gene on AML cells, we investigated the biologic and molecular mechanism associated with AML1-ETO expressed in t(8;21) AML.</p><p><b>METHODS</b>Qualitative polymerase chain reaction (PCR), quantitative reverse transcription PCR (RT-PCR), and Western blotting analysis were used to observe the mRNA and protein expression levels of EYA4 in cell lines. Different plasmids (including mutant plasmids) of dual luciferase reporter vector were built to study the binding status of AML1-ETO to the promoter region of EYA4. Chromatin immunoprecipitation assay was used to study the epigenetic silencing mechanism of EYA4. Bisulfite sequencing was applied to detect the methylation status in EYA4 promoter region. The influence of EYA4 gene in the cell proliferation, apoptosis, and cell clone-forming ability was detected by the technique of Cell Counting Kit-8, flow cytometry, and clonogenic assay.</p><p><b>RESULTS</b>EYA4 gene was hypermethylated in AML1-ETO+ patients and its expression was down-regulated by 6-fold in Kasumi-1 and SKNO-1 cells, compared to HL-60 and SKNO-1-siA/E cells, respectively. We demonstrated that AML1-ETO triggered the epigenetic silencing of EYA4 gene by binding at AML1-binding sites and recruiting histone deacetylase 1 and DNA methyltransferases. Enhanced EYA4 expression levels inhibited cellular proliferation and suppressed cell colony formation in AML1-ETO+ cell lines. We also found EYA4 transfection increased apoptosis of Kasumi-1 and SKNO-1 cells by 1.6-fold and 1.4-fold compared to negative control, respectively.</p><p><b>CONCLUSIONS</b>Our study identified EYA4 gene as targets for AML1-ETO and indicated it as a novel tumor suppressor gene. In addition, we provided evidence that EYA4 gene might be a novel therapeutic target and a potential candidate for treating AML1-ETO+ t (8;21) AML.</p>


Subject(s)
Apoptosis , Genetics , Physiology , Blotting, Western , Cell Line, Tumor , Cell Proliferation , Genetics , Physiology , Chromatin Immunoprecipitation , Core Binding Factor Alpha 2 Subunit , Genetics , Metabolism , DNA Methylation , Genetics , Epigenesis, Genetic , Genetics , Gene Silencing , HL-60 Cells , Humans , Leukemia, Myeloid, Acute , Genetics , Metabolism , Pathology , Oncogene Proteins, Fusion , Genetics , Metabolism , RNA, Small Interfering , Genetics , RUNX1 Translocation Partner 1 Protein , Radioimmunoprecipitation Assay , Trans-Activators , Genetics , Metabolism
15.
Article in Chinese | WPRIM | ID: wpr-300838

ABSTRACT

To investigate the effect ofgene down-regulation on early hematopoietic development of zebrafish.Phosphorodiamidate morpholino oligomer (PMO) technology was used to downregulategene expression in Zebrafish. Zebrafish embryos injected phosphorodiamidate morpholino antisense oligonucleotide ofgene mRNA by microinjection at unicellular stage were taken as the experimental group, and those injected meaningless phosphorodiamidate morpholino antisense oligonucleotide were taken as the control. The embryos were collected at 18, 24, 30 and 36 hpf after the fertilization. The real-time fluorescent quantitative PCR (RT-PCR) and whole embryohybridization methods were used to detect the expression of myeloid hematopoietic transcription factorand erythroid hematopoietic transcription factorin zebrafish.RT-PCR showed that the expressions ofanddecreased in the experimental group compared with the control group (all<0.05). Whole embryohybridization showed that the blue-black positive hybridization signals ofandin experimental group were shallow than those in the control group.Myeloid hematopoietic and erythroid hematopoietic of zebrafish are blocked with the downregulation ofgene.


Subject(s)
Animals , Down-Regulation , Genetics , Embryo, Nonmammalian , GATA1 Transcription Factor , Genetics , Metabolism , Gene Knockdown Techniques , Hematopoiesis , In Situ Hybridization , Lamin Type A , Genetics , Physiology , Proto-Oncogene Proteins , Genetics , Metabolism , Trans-Activators , Genetics , Metabolism , Zebrafish , Embryology , Genetics
16.
Article in Chinese | WPRIM | ID: wpr-264011

ABSTRACT

<p><b>OBJECTIVE</b>To investigate whether phenotypic modulation of bladder smooth muscle occurs in diabetic rats.</p><p><b>METHODS</b>Thirty-two male SD rats were randomly assigned into diabetic group and control group. Diabetic rat models were established by a single intraperitoneal injection of streptozotocin (60 mg/kg). Nine weeks later, the bladder tissues of the rats were examined for structural changes using HE and Masson's trichrome staining , and the expressions of myocardin, α-SMA, and SMMHC in bladder smooth muscles were detected with RT-PCR and Western blotting.</p><p><b>RESULTS</b>Compared with the control group, the diabetic rats showed obvious polydipsia and polyuria with significantly increased collagenous fibers and lowered expressions of myocardin, α-SMA, and SMMHC in the bladder tissue (P<0.05).</p><p><b>CONCLUSION</b>s In rats at 9 weeks after diabetic model establishment, phenotypic transition of the bladder smooth muscles occurs to cause bladder contractile dysfunction, which may play an important role in the pathology of diabetic bladder dysfunction.</p>


Subject(s)
Actins , Metabolism , Animals , Diabetes Mellitus, Experimental , Male , Muscle Contraction , Muscle, Smooth , Myosin Heavy Chains , Metabolism , Nuclear Proteins , Metabolism , Phenotype , Rats , Rats, Sprague-Dawley , Streptozocin , Trans-Activators , Metabolism , Urinary Bladder
17.
Chinese Journal of Hepatology ; (12): 152-156, 2016.
Article in Chinese | WPRIM | ID: wpr-314644

ABSTRACT

Hepatocellular carcinoma (HCC) is one of the most common cancer worldwide. Most of the HCC occur in developing countries. Chronic hepatitis B virus (HBV) infection is an important risk factor for HCC development. HBV induces immune-mediated chronic hepatitis, liver injury, regeneration and scar forming responses, leading to an inflammatory, fibrotic and immune deficient microenvironment. HBV may integrate into host genome, inducing genetic abnormality and altering the expression of HCC-related genes. HBV also expresses active proteins such as X (HBx) and S proteins, which may trans-activate HCC-related proteins expression, interact with intracellular specific proteins, activate a variety of signaling pathways, and induce aberrant epigenetic modifications. HBV mutation also has impact on HBV related HCC development.


Subject(s)
Carcinoma, Hepatocellular , Pathology , Virology , Epigenesis, Genetic , Hepatitis B virus , Hepatitis B, Chronic , Pathology , Humans , Liver Neoplasms , Pathology , Virology , Mutation , Signal Transduction , Trans-Activators
18.
Article in English | WPRIM | ID: wpr-84906

ABSTRACT

This study aimed to validate the high yield and soluble expression of proteins carrying the transactivator of transcription (Tat) peptide tag, and further explored the potential mechanism by which the Tat tag increases expression. Escherichia coli superoxide dismutase (SOD) proteins, including SodA, SodB and SodC, were selected for analysis. As expected, the yields and the solubility of Tat-tagged proteins were higher than those of Tat-free proteins, and similar results were observed for the total SOD enzyme activity. Bacterial cells that overexpressed Tat-tagged proteins exhibited increased anti-paraquat activity compared with those expressing Tat-free proteins that manifested as SodA>SodC>SodB. When compared with an MG1655 wild-type strain, the growth of a ΔSodA mutant strain was found to be inhibited after paraquat treatment; the growth of ΔSodB and ΔSodC mutant strains was also slightly inhibited. The mRNA transcript level of genes encoding Tat-tagged proteins was higher than that of genes encoding Tat-free proteins. Furthermore, the α-helix and turn of Tat-tagged proteins were higher than those of Tat-free proteins, but the β-sheet and random coil content was lower. These results indicated that the incorporation of the Tat core peptide as a significant basic membrane transduction peptide in fusion proteins could increase mRNA transcripts and promote the high yield and soluble expression of heterologous proteins in E. coli.


Subject(s)
Escherichia coli , Escherichia , HIV-1 , Membranes , Paraquat , RNA, Messenger , Solubility , Superoxide Dismutase , Superoxides , Trans-Activators
19.
Rev. latinoam. enferm. (Online) ; 23(4): 635-641, July-Aug. 2015. tab
Article in English | LILACS, BDENF | ID: lil-761706

ABSTRACT

AbstractObjective: to validate the content of the prevention protocol for early sepsis caused by Streptococcus agalactiaein newborns.Method: a transversal, descriptive and methodological study, with a quantitative approach. The sample was composed of 15 judges, 8 obstetricians and 7 pediatricians. The validation occurred through the assessment of the content of the protocol by the judges that received the instrument for data collection - checklist - which contained 7 items that represent the requisites to be met by the protocol. The validation of the content was achieved by applying the Content Validity Index.Result: in the judging process, all the items that represented requirements considered by the protocol obtained concordance within the established level (Content Validity Index > 0.75). Of 7 items, 6 have obtained full concordance (Content Validity Index 1.0) and the feasibility item obtained a Content Validity Index of 0.93. The global assessment of the instruments obtained a Content Validity Index of 0.99.Conclusion: the validation of content that was done was an efficient tool for the adjustment of the protocol, according to the judgment of experienced professionals, which demonstrates the importance of conducting a previous validation of the instruments. It is expected that this study will serve as an incentive for the adoption of universal tracking by other institutions through validated protocols.


ResumoObjetivo:validar o conteúdo do protocolo de prevenção da sepse precoce porStreptococcus agalactiaeem recém-nascidos.Método:estudo transversal, descritivo, do tipo metodológico, com abordagem quantitativa. A amostra foi composta por 15 juízes, oito médicos obstetras e sete pediatras. A validação ocorreu por intermédio da avaliação de conteúdo do protocolo pelos juízes, os quais receberam o instrumento de coleta de dados - checklist - contendo sete itens, que representam requisitos a serem contemplados no protocolo. A validação de conteúdo foi atingida mediante aplicação do Índice de Validade de Conteúdo.Resultado:no processo de julgamento, todos os itens que representam requisitos contemplados no protocolo obtiveram concordância dentro do nível estabelecido (Índice de Validade de Conteúdo >0,75). Dos sete itens, seis obtiveram concordância total, (Índice de Validade de Conteúdo 1.0) e o item exequibilidade obteve Índice de Validade de Conteúdo de 0,93. A avaliação global dos instrumentos obteve Índice de Validade de Conteúdo de 0,99.Conclusão:a validação de conteúdo realizada foi ferramenta eficaz para adequação do protocolo, de acordo com o julgamento de profissionais experientes, demonstrando a importância em se realizar validação prévia de instrumentos. Espera-se que, este estudo incentive a adoção do rastreio universal por outras instituições, mediante protocolos validados.


ResumenObjetivo:validar el contenido del protocolo de prevención de la sepsis precoz porStreptococcus agalactiaeen recién nacidos.Método:estudio transversal, descriptivo, del tipo metodológico, con un enfoque cuantitativo. La muestra fue conformada por 15 jueces, ocho obstetras y siete pediatras. La validación se dio a través de la evaluación de contenido del protocolo por los jueces, los cuales recibieron el instrumento de recolección de datos - checklist - conteniendo siete ítems, que representan los requisitos para ser incluidos en el protocolo. La validación de contenido se logró a través de la aplicación del Índice de Validez de Contenido.Resultado:en el proceso de evaluación, todos los ítems que representan los requisitos contemplados en el protocolo obtuvieron una concordancia dentro del nivel establecido (Índice de Validez de Contenido > 0,75). De los siete ítems, seis obtuvieron una concordancia total (Índice de Validez de Contenido 1,0), y el ítem viabilidad obtuvo un Índice de Validez de Contenido de 0,93. La evaluación global de los instrumentos obtuvo un Índice de Validez de Contenido de 0,99.Conclusión:la validación de contenido realizada fue una herramienta eficaz para la adecuación del protocolo, según la evaluación de profesionales expertos, demostrando así la importancia de realizar la validación previa de los instrumentos. Se espera que este estudio fomente la adopción del cribado (screening) universal por otras instituciones, mediante protocolos validados.


Subject(s)
Humans , Animals , Arteries/transplantation , Graft Survival , Nuclear Proteins , Organ Transplantation , Trans-Activators , Transplantation Tolerance/genetics , Animals, Genetically Modified , Graft Survival/genetics , Graft Survival/immunology , Heterografts , Nuclear Proteins/genetics , Nuclear Proteins/immunology , Papio , Swine , Trans-Activators/genetics , Trans-Activators/immunology
20.
Rev. argent. microbiol ; 47(2): 88-94, June 2015. tab
Article in Spanish | LILACS | ID: lil-757146

ABSTRACT

En la provincia del Chaco, el agua subterránea representa una fuente alternativa, y muchas veces única, para el consumo humano; esta es utilizada en el 14 % de los hogares. A pesar de que se reconoce el riesgo de la exposición al agua contaminada, la prevalencia de los diferentes patotipos de Escherichia coli en ambientes acuáticos no ha sido bien caracterizada. E. coli enteroagregativo (ECEA) es un patógeno emergente cuya importancia en la salud pública mundial se incrementó y quedó claramente establecida en los últimos años. El objetivo del presente trabajo fue detectar la presencia de ECEA típico mediante el reconocimiento de los factores de virulencia aap, AA probe y aggR por reacción en cadena de la polimerasa, en fuentes de agua subterráneas de la provincia del Chaco. Se identificó E. coli en 36 (38,7 %) de las 93 muestras estudiadas, provenientes de diferentes localidades. De esos 36 aislamientos, se identificaron 6 (16,7 %) portadores de los genes de ECEA, lo que representa una prevalencia del 6,4 % considerando las 93 fuentes de agua subterránea estudiadas. De esos 6 aislamientos, 3 eran portadores del gen aap, 2 del gen AA probe y uno de la combinación aggR/aap. El presente trabajo representa el primer aporte en el estudio de la presencia y distribución de genes de virulencia de ECEA en fuentes de agua subterránea de la región.


Groundwater is an important source of drinking water for many communities in Northern Argentina; particularly, in the province of Chaco, where about 14 % of households use this natural resource. Enteroaggregative Escherichia coli is an emerging pathogen whose global importance in public health has increased in recent years. Despite the significant risk of disease linked to contaminated water exposure, the prevalence of E. coli pathotypes in aquatic environments is still not so well defined. The aim of the present study was to detect the presence of typical enteroaggregative E. coli through the recognition of its virulence factors aap, AA probe and aggR by molecular techniques. A total of 93 water samples from different small communities of Chaco were analyzed. E. coli was identified in 36 (38.7 %) of the tested samples. Six strains isolated from different samples harbored the studied genes. Of these 6 isolates, 3 carried the aap gene, 2 the AA probe and the last one the combination of aap/aggR genes. The prevalence of E. coli isolates harboring enteroaggregative virulence genes in groundwater sources was 6.4 %. This work represents the first contribution to the study of the presence and distribution of virulence genes of EAEC in groundwater sources in this region of Argentina.


Subject(s)
Escherichia coli Proteins/genetics , Escherichia coli/genetics , Genes, Bacterial , Groundwater/microbiology , Trans-Activators/genetics , Water Pollution , Argentina , Escherichia coli Proteins/physiology , Escherichia coli/isolation & purification , Escherichia coli/pathogenicity , Trans-Activators/physiology , Virulence/genetics , Water Supply
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