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1.
Int Immunopharmacol ; 84: 106541, 2020 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-32402950

RESUMO

OBJECTIVE: To illustrate the potential effects and mechanism of extracellular vesicles derived from mesenchymal stem cells (MSC-EVs) on fibrosis in sclerodermatous chronic graft-versus-host-disease (cGVHD) models after allogeneic hematopoietic stem cell transplantation. METHODS: We first observed the therapeutic effects of MSC-EVs on a minor histocompatibility haploidentical model of sclerodermatous cGVHD and the function of MSC-EVs on skin fibrosis and macrophage activation and the related pro-fibrosis protein. Additionally, we observed the effects of MSC-EVs on B cells, the T follicular helper cell (TFH) and germinal center B cell (GC B cells) interaction and the ratio of B cell activation factor (BAFF) to B cells in vivo. RESULTS: MSC-EVs treatment could alleviate the cGVHD scores and fibrosis of skin in sclerodermatous cGVHD mice, and this was associated with a reduction macrophage percentage in the skin and spleen, and a reduction in macrophage infiltration and TGF-ß and smad2 production in the skin. Additionally, MSC-EVs influence B cells immune response by blocking the TFH/GC B cells interaction and reducing the ratio of BAFF to B cells in vivo. CONCLUSION: MSC-EVs prevent the fibrosis of sclerodermatous cGVHD mouse model by suppressing the activation of macrophages and B cells immune response.

2.
Clin Appl Thromb Hemost ; 26: 1076029619892684, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32088973

RESUMO

CONCLUSION: von Willebrand factor is a useful predictor and prognostic measure for TA-TMA, which may help clinicians identify and manage this life-threatening disease earlier.

3.
Cytokine ; 126: 154868, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31629110

RESUMO

Lung cancer is a common malignant disease, nearly 2.09 million new patients occurred last year. Approximately 85% of the patients are classified as non-small-cell lung cancer (NSCLC). It is therefore important to identify new diagnostic and prognostic biomarkers for the early detection of this disease. The presented study identifies biomarkers in the serum of NSCLC patients. The expression of 274 cytokines was measured by a novel antibody array methodology and ELISA was applied to validate the array results. The levels of MIP-1 α, IL-8, MIP-1 ß, Resistin, GDF-15, HGF, CA125, FLRG, VCAM-1, DKK-3, sTNF-R1, CTACK, Acrp30, CXCL-16 and LYVE-1 were significantly higher in serum from NSCLC patients, while the level of TIMP-2 and IGFBP-6 were lower. More importantly, the validation supported the result of the antibody array. The result of the antibody array indicates that these cytokines might be novel auxiliary biomarkers in the diagnosis and prognosis of NSCLC.

4.
J Hematol Oncol ; 12(1): 117, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31747962

RESUMO

The original article [1] contains an error in authorship whereby author, Robert Weinkove's name is mistakenly inverted. The configuration noted in this Correction article should be considered instead along with author's updated affiliation.

5.
BMC Infect Dis ; 19(1): 631, 2019 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-31315565

RESUMO

BACKGROUND: Candida arthritis is extremely rare and also represents a major challenge of diagnosis and treatment. Here we reported a rare case of recurrent arthritis caused by Candida parapsilosis. CASE PRESENTATION: A 56-year-old Chinese male suffered from recurrent pain and swelling in his right knee after several times of "small needle-knife" acupuncture and corticosteroid injection of the joint. Candida parapsilosis was cultured in his synovial fluid and identified by sequencing of its Internal Transcribed Spacer (ITS) gene. Here we present the radiological characteristics, arthroscopic pictures, and synovium pathology of this patient. Also, blood test and chemical analysis of his synovial fluid were listed as well as the ITS sequence of this Candida species identified. The patient underwent thorough arthroscopic debridement and then set on fluconazole 400 mg daily for 12 months. His symptoms resolved and no relapse was observed on the last follow-up. Additionally, a brief but comprehensive review of C. parapsilosis arthritis episodes from past to now were studied. CONCLUSION: With the detailed clinical information reported in this case and our literature review, we hope they would add to our knowledge of C. parapsilosis arthritis - its clinical settings, laboratory features, radiological characteristics, arthroscopic findings and experience of management.


Assuntos
Artrite/microbiologia , Candida parapsilosis/patogenicidade , Candidíase/tratamento farmacológico , Antifúngicos/uso terapêutico , Artrite/tratamento farmacológico , Artrite/cirurgia , Candida parapsilosis/isolamento & purificação , Desbridamento , Fluconazol/uso terapêutico , Humanos , Joelho/microbiologia , Joelho/patologia , Masculino , Pessoa de Meia-Idade , Líquido Sinovial/microbiologia
6.
Int Immunopharmacol ; 74: 105663, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31200338

RESUMO

Extracellular vesicles (EVs) secreted by mesenchymal stem cells (MSC-EVs) are taken more seriously as immunomodulatory and anti-inflammatory agents. We studied the therapeutic effects of MSC-EVs on allergic contact dermatitis (ACD), a typical T cell-mediated disorder. A contact hypersensitivity (CHS) mouse model for ACD was established and treated by intravenous MSC-EVs injection. We found that human umbilical cord MSC-EVs could significantly prevent the pathology of CHS, including reduced ear swelling and leukocyte infiltration. Injection of MSC-EVs significantly inhibited CD8+IFN-γ+ cytotoxic T (Tc1) cells and CD4+IFN-γ+ type 1 helper T (Th1) cells, and reduced the level of pro-inflammatory Tumor Necrosis Factor-alpha (TNF-α) and interferon gamma (IFN-γ), and induced CD4+CD25+Foxp3+ regulatory T cells (Tregs) and the level of anti-inflammatory IL-10. In vitro, MSC-EVs also suppressed Tc1 and Th1 cells and induced Tregs and the related cytokines, further indicating the immune regulatory role of MSC-EVs. Interestingly, PKH26-labeled MSC-EVs were found to be directly internalized by CD3+ T cells, resulting in reduced signal transducer and activator of transcription 1 (STAT1) protein levels in vitro. In summary, MSC-EVs can prevent the onset of CHS by inhibiting Tc1 and Th1 immune responses and inducing the Tregs phenotype in vivo and in vitro. The mechanism by which MSC-EVs influence CD3+ T cells might partially involve targeting STAT1 in vitro. Therefore, MSC-EVs are ideal candidates for cell-free immunomodulatory therapy for T cell-mediated diseases such as ACD.


Assuntos
Dermatite de Contato/imunologia , Vesículas Extracelulares/imunologia , Leucócitos Mononucleares/imunologia , Células-Tronco Mesenquimais/imunologia , Linfócitos T/imunologia , Animais , Citocinas/genética , Citocinas/imunologia , Humanos , Masculino , Camundongos Endogâmicos BALB C
7.
J Hematol Oncol ; 11(1): 25, 2018 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-29458388

RESUMO

BACKGROUND: Anti-CD19 chimeric antigen receptor (CAR) T cells have shown promise in the treatment of B cell acute lymphocytic leukemia (B-ALL). However, its efficacy in B-ALL patients with extramedullary involvement is limited due to poor responses and neurotoxicity. Here, we utilized a third generation of CAR T cell vector, which contains the Toll/interleukin-1 receptor (ITR) domain of Toll-like receptor 2 (TLR2), to generate 1928zT2 T cells targeting CD19, and evaluated the efficacy of 1928zT2 T cells in relapse or refractory B-ALL patients with extramedullary involvement. METHODS: 1928zT2 T cells were generated by 19-28z-TLR2 lentiviral vector transfection into primary human T lymphocytes. The anti-leukemia effect of 1928zT2 T cells were determined by killing assays and in xenografts. Three patients diagnosed as relapse or refractory ALL with extramedullary involvement were infused with 1928zT2 T cells, and the clinical responses were evaluated by BM smear, B-ultrasonography, PET/CT, histology, flow cytometry, qPCR, ELISA, and luminex assay. RESULTS: 1928zT2 T cells exhibited enhanced effector function against CD19+ leukemic cells in vitro and in a xenograft model of human extramedullary leukemia. Notably, the 1928zT2 T cells eradicated extramedullary leukemia and induced complete remission in the three relapse and refractory ALL patients without serious adverse effects. 1928zT2 T cells expanded robustly in the circulation of these three patients and were detected in the cerebrospinal fluid of patient 3. These three patients experienced cytokine release syndrome (CRS) with grade 2 or 3, which remitted spontaneously or after tocilizumab treatment. None of the three patients suffered neurotoxicity or needed further intensive care. CONCLUSIONS: Our results demonstrate that 1928zT2 T cells with TLR2 incorporation augment anti-leukemic effects, particularly for eradicating extramedullary leukemia cells, and suggest that the infusion of 1928zT2 T cells is an encouraging treatment for relapsed/refractory ALL patients with extramedullary involvement. TRIAL REGISTRATION: ClinicalTrials.gov identifier NCT02822326 . Date of registration: July 4, 2016.


Assuntos
Imunoterapia Adotiva/métodos , Recidiva Local de Neoplasia/terapia , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Receptores de Antígenos Quiméricos/imunologia , Linfócitos T/imunologia , Receptor 2 Toll-Like/imunologia , Adolescente , Adulto , Animais , Antígenos CD19/imunologia , Células Cultivadas , Feminino , Humanos , Masculino , Camundongos , Recidiva Local de Neoplasia/imunologia , Recidiva Local de Neoplasia/patologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/imunologia , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Receptores de Antígenos Quiméricos/genética , Linfócitos T/patologia , Linfócitos T/transplante , Receptor 2 Toll-Like/genética , Transfecção , Adulto Jovem
8.
Sci Rep ; 8(1): 2855, 2018 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-29434267

RESUMO

To examine the early changes of articular cartilage and subchondral bone in the DMM mouse model of osteoarthritis, mice were subjected to DMM or SHAM surgery and sacrificed at 2-, 5- and 10-week post-surgery. Catwalk gait analyses, Micro-Computed Tomography, Toluidine Blue, Picrosirius Red and Tartrate-Resistant Acid Phosphatase (TRAP) staining were used to investigate gait patterns, joint morphology, subchondral bone, cartilage, collagen organization and osteoclasts activity, respectively. Results showed OA progressed over 10-week time-course. Gait disparity occurred only at 10-week post-surgery. Osteophyte formed at 2-week post-surgery. BMDs of DMM showed no statistical differences comparing to SHAM at 2 weeks, but BV/TV is much higher in DMM mice. Increased BMD was clearly found at 5- and 10-week post-surgery in DMM mice. TRAP staining showed increased osteoclast activity at the site of osteophyte formation of DMM joints at 5- and 10-week time points. These results showed that subchondral bone turnover might occurred earlier than 2 weeks in this mouse DMM model. Gait disparity only occurred at later stage of OA in DMM mice. Notably, patella dislocation could occur in some of the DMM mice and cause a different pattern of OA in affected knee.


Assuntos
Cartilagem Articular/diagnóstico por imagem , Osteoartrite do Joelho/diagnóstico por imagem , Osteoclastos/patologia , Osteófito/diagnóstico por imagem , Animais , Cartilagem Articular/patologia , Modelos Animais de Doenças , Análise da Marcha , Humanos , Meniscos Tibiais/cirurgia , Camundongos , Osteoartrite do Joelho/etiologia , Osteoartrite do Joelho/patologia , Osteófito/patologia , Fatores de Tempo , Microtomografia por Raio-X
9.
Clin Chim Acta ; 440: 57-63, 2015 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-25447698

RESUMO

BACKGROUND: HE4, a novel tumor marker for detecting ovarian cancer, has been recently applied to clinical practice. However, the comprehensive evaluation of HE4 combined with other markers is still missing. We evaluated an optimal mode of HE4 employment for differential diagnosis of benign and malignant pelvic masses. METHODS: Serum HE4, CA125, CA153, CA199, CA211 and CA724 were measured from 232 patients with pelvic messes (100 malignant masses, 132 benign diseases), and the risk of ovarian malignancy algorithm (ROMA) was also calculated. Receiver operating characteristic curves (ROC), the area under the curve (AUC), sensitivity and specificity were estimated. RESULTS: The combination of HE4 and CA125 (AUC of 0.963, sensitivity of 96.6%, specificity of 65.7%) provided the best differential power in diagnosing ovarian cancer. ROMA performed better in the diagnosis of pelvic masses (AUC of 0.917, sensitivity of 82.0%, specificity of 78.8%) and uterine cancer (AUC of 0.838, sensitivity of 82.0%, specificity of 60.0%) compared with applying HE4 and CA125 individually. CONCLUSION: The optimal cut-off values (CA125: 93.2U/ml, HE4: 87.6 pmol/l, ROMA: 18.1% for pre- and 31.5% for postmenopausal women), simultaneous use of CA125 and HE4 complemented by ROMA showed better performance than the traditional detection modes for differential diagnosis of ovarian cancer. We also observed that ROMA added more accuracy for differentiating the benign and malignant pelvic masses and auxiliary diagnosis of uterine cancer.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias Ovarianas/diagnóstico , Neoplasias Pélvicas/diagnóstico , Adulto , Idoso , Algoritmos , Antígenos Glicosídicos Associados a Tumores/sangue , Área Sob a Curva , Antígeno Ca-125/sangue , Diagnóstico Diferencial , Neoplasias do Endométrio/sangue , Neoplasias do Endométrio/diagnóstico , Neoplasias do Endométrio/patologia , Feminino , Humanos , Proteínas de Membrana/sangue , Pessoa de Meia-Idade , Neoplasias Ovarianas/sangue , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/cirurgia , Neoplasias Pélvicas/sangue , Neoplasias Pélvicas/patologia , Neoplasias Pélvicas/cirurgia , Proteínas/análise , Sensibilidade e Especificidade , Neoplasias Uterinas/sangue , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/patologia , Neoplasias Uterinas/cirurgia , Adulto Jovem
10.
Parasitol Res ; 113(12): 4543-53, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25300416

RESUMO

Globally, 15-20 million people are infected with Clonorchis sinensis (C. sinensis) which results in clonorchiasis. In China, clonorchiasis is considered to be one of the fastest-growing food-borne parasitic diseases. That more key molecules of C. sinensis are characterized will be helpful to understand biology and pathogenesis of the carcinogenic liver fluke. Glyceraldehyde-3-phosphate dehydrogenases (GAPDHs) from many species have functions other than their catalytic role in glycolysis. In the present study, we analyzed the sequence and structure of GAPDH from C. sinensis (CsGAPDH) by using bioinformatics tools and obtained its recombinant protein by prokaryotic expression system, to learn its expression profiles and molecular property. CsGAPDH could bind to human intrahepatic biliary epithelial cell in vivo and in vitro by the method of immunofluorescence assays. CsGAPDH also disturbed in lumen of biliary tract near to the parasite in the liver of infected rat. Western blotting analysis together with immunofluorescence assay indicated that CsGAPDH was a component of excretory/secretory proteins (CsESPs) and a surface-localized protein of C. sinensis. Quantitative real-time PCR (Q-PCR) and Western blotting demonstrated that CsGAPDHs are expressed at the life stages of adult worm, metacercaria, and egg, but the expression levels were different from each other. Recombinant CsGAPDH (rCsGAPDH) was confirmed to have the capacity to catalyze the conversion of glyceraldehyde 3-phosphate to D-glycerate 1,3-bisphosphate which was inhibited by AMP in a dose-dependent manner. In addition, rCsGAPDH was able to interact with human plasminogen in a dose-dependent manner by ELISA. The interaction could be inhibited by lysine. The plasminogen binding capacity of rCsGAPDH along with the distribution of CsGAPDH in vivo and in the liver of C. sinensis-infected rat hinted that surface-localized CsGAPDH might play an important role in host invasion of the worm besides its glycolytic activity. Our work will be a cornerstone for getting more messages about CsGAPDH and its role in biology and parasitism of C. sinensis.


Assuntos
Clonorquíase/parasitologia , Clonorchis sinensis/enzimologia , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Plasminogênio/metabolismo , Sequência de Aminoácidos , Animais , Western Blotting , China , Clonorquíase/enzimologia , Clonorchis sinensis/genética , Biologia Computacional , Regulação Enzimológica da Expressão Gênica , Gliceraldeído-3-Fosfato Desidrogenases/química , Gliceraldeído-3-Fosfato Desidrogenases/genética , Proteínas de Helminto/isolamento & purificação , Humanos , Fígado/enzimologia , Fígado/parasitologia , Metacercárias/enzimologia , Camundongos , Camundongos Endogâmicos BALB C , RNA de Helmintos/isolamento & purificação , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/genética
11.
Exp Ther Med ; 8(1): 213-218, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24944624

RESUMO

Interleukin-21 (IL-21) is a pleiotropic cytokine linking innate and adaptive immune responses, which has been reported to play a key role in multiple autoimmune diseases. The aim of the present case-control study was to investigate the genetic association between single nucleotide polymorphisms (SNPs) of rs907715 within the IL-21 gene and Graves' disease (GD) in a Southern Chinese population. A total of 211 patients with GD and 212 control subjects were recruited for the study. IL-21 gene rs907715 polymorphisms were detected by direct DNA sequencing. The results indicated that the frequencies of the GG genotype and the G allele in GD patients were significantly increased when compared with the frequencies in the controls (P=6.7×10-3 and P=2.0×10-5, respectively). In addition, the frequency of the AA genotype was much lower in the patient group when compared with the control group (16.6 vs. 34.0%; P=4.0×10-5). Furthermore, the G allele of rs907715 was associated with relapse in GD patients. These observations indicated that polymorphisms of IL-21/rs907715 may affect the susceptibility to GD in a Southern Chinese population. The G allele was significantly associated with an increased risk of GD development, whereas the A allele may lower the susceptibility to GD.

12.
Parasitol Res ; 113(5): 1713-23, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24578258

RESUMO

This is the first report of a novel protein from Clonorchis sinensis (C. sinensis), serine/threonine protein kinase 17A (CsSTK17A), which belongs to a member of the death-associated protein kinase (DAPK) family known to regulate diverse biological processes. The full-length sequence encoding CsSTK17A was isolated from C. sinensis adult cDNA plasmid library. Two transcribed isoforms of the gene were identified from the genome of C. sinensis. CsSTK17A contains a kinase domain at the N-terminus that shares a degree of conservation with the DAPK families. Besides, the catalytic domain contains 11 subdomains conserved among STKs and shares the highest identity with STK from Schistosoma mansoni (55.9%). Three-dimensional structure of CsSTK17A displays the canonical STK fold, including the helix C, P-loop, and the activation loop. We obtained recombinant CsSTK17A (rCsSTK17A) and anti-rCsSTK17A IgG. The rCsSTK17A could be probed by anti-rCsSTK17A rat serum, C. sinensis-infected rat serum and the sera from rats immunized with C. sinensis excretory-secretory products, indicating that it is a circulating antigen possessing a strong immunocompetence. Moreover, quantitative RT-PCR and western blotting analyses revealed that CsSTK17A exhibited the highest mRNA and protein expression level in eggs, followed by metacercariae and adult worms. Intriguingly, in the immunolocalization assay, CsSTK17A was intensively localized to the operculum region of eggs in uterus, as well as the vitelline gland of both adult worm and metacercaria, implying that the protein was associated with the reproduction and development of C. sinensis. Overall, these fundamental studies might contribute to further researches on signaling systems of the parasite.


Assuntos
Clonorchis sinensis/enzimologia , Proteínas de Helminto/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Sequência de Aminoácidos , Animais , Clonagem Molecular , Clonorchis sinensis/genética , Biblioteca Gênica , Proteínas de Helminto/genética , Modelos Moleculares , Dados de Sequência Molecular , Estrutura Terciária de Proteína , Proteínas Serina-Treonina Quinases/genética , Ratos , Ratos Sprague-Dawley , Análise de Sequência de DNA
13.
Parasitol Res ; 113(4): 1545-54, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24535733

RESUMO

The vacuolar ATPase enzyme complex (V-ATPase) pumps protons across membranes, energized by hydrolysis of ATP. Extensive investigations on structural and biochemical features of these molecules have implied their importance in the physiological process. In this study, a full-length sequence encoding a vacuolar ATP synthase subunit ε-like protein of Clonorchis sinensis (CsATP-ε) was isolated from our cDNA library. The hypothetical 226 amino acid sequence shared 76% identity with ATP-ε proteins of Schistosoma japonicum and above 55% identity with ATP-ε proteins from human and other eukaryotes. Characteristic Asp140 amino acid residues and seven B-cell epitopes were predicted in this sequence. The complete coding sequence of the gene was expressed in Escherichia coli. Recombinant CsATP-ε (rCsATP-ε) protein could be probed by anti-rCsATP-ε rat serum and C.sinensis-infected human serum in Western blotting experiment, indicating that it is an antigen of strong antigenicity. The high level of antibody titers (1:204,800) showed that CsATP-ε has a powerful immunogenicity. Both the increased level and the change trend of IgG1/IgG2a subtypes in serum showed that the rCsATP-ε can induce strong combined Th1/Th2 immune responses in rats and stimulate the immune response changes to the dominant Th2 from Th1 along with long time infection. The results of immunoblot and immunolocalization demonstrated that CsATP-ε was consecutively expressed at various developmental stages of the parasite, which was supported by real-time PCR analysis. In immunohistochemistry, CsATP-ε was localized on the intestine, vitellarium, and testicle of an adult worm and excretory bladder of metacercaria, implying that CsATP-ε may relate to energy intake and metabolism. This fundamental study would contribute to further researches that are related to growth and development and immunomodulation of C. sinensis.


Assuntos
Clonorchis sinensis/enzimologia , Etenoadenosina Trifosfato/imunologia , ATPases Vacuolares Próton-Translocadoras/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Anti-Helmínticos/sangue , Clonagem Molecular , Clonorchis sinensis/genética , Clonorchis sinensis/imunologia , Epitopos de Linfócito B/imunologia , Humanos , Imunidade Celular , Imunoglobulina G/sangue , Dados de Sequência Molecular , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia
14.
Zhonghua Xue Ye Xue Za Zhi ; 34(7): 561-5, 2013 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-23906446

RESUMO

OBJECTIVE: To understand the predictive value of early monitoring BCR-ABL transcripts in patients with chronic myeloid leukemia (CML) after treatment with tyrosine kinase inhibitor (TKI), and to provides the information for early assessment of prognosis and treatment options. METHODS: BCR-ABL transcripts of 53 CML patients before and after TKI treatment were detected by using real-time quantitative RT-PCR. The relationship between BCR-ABL transcripts level after TKI treatment for 3 months and the later molecular response, progression and mutation was analyzed. RESULTS: The median values of BCR-ABL transcripts in peripheral blood samples from 30 newly diagnosed patients were 43.99%, which was used as a baseline of BCR-ABL transcripts for molecular response evaluation. Of 53 patients, 31 (58.49%) had a BCR-ABL mRNA ≤ 4.40% (reduced more than 1 log) and 22 (41.51%) greater than 4.40% (reduced to less than 1 log) after 3 months of TKI treatment. The former 31 patients had a significantly higher 18-months cumulative incidence of major molecular response (MMR) (90.32% vs 18.18%, P=0.000) and 3-year cumulative incidence of complete molecular response (CMR) (48.39% vs 0, P=0.000) compared with the latter 22 patients. The lower BCR-ABL level was, the earlier MMR reached. The proportion of patients with a mutation in group of BCR-ABL mRNA>4.40% was significantly higher than that of BCR-ABL mRNA ≤ 4.40% (22.73% vs 0, P=0.021). The incidence of progression increased in group of BCR-ABL mRNA>4.40%, but the difference was not statistically significant (P=0.052). CONCLUSION: It is important for the prognosis evaluation of the patients to monitor the level of BCR-ABL transcripts at 3 months after TKI treatment, which might help to early optimization of treatment and to improve curative effect of CML patients.


Assuntos
Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Inibidores de Proteínas Quinases/uso terapêutico , Adulto , Idoso , Feminino , Proteínas de Fusão bcr-abl/sangue , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , RNA Mensageiro/genética , Resultado do Tratamento , Adulto Jovem
15.
PLoS One ; 8(1): e54732, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23382950

RESUMO

Clonorchis sinensis (C. sinensis), an important food-borne parasite that inhabits the intrahepatic bile duct and causes clonorchiasis, is of interest to both the public health field and the scientific research community. To learn more about the migration, parasitism and pathogenesis of C. sinensis at the molecular level, the present study developed an upgraded genomic assembly and annotation by sequencing paired-end and mate-paired libraries. We also performed transcriptome sequence analyses on multiple C. sinensis tissues (sucker, muscle, ovary and testis). Genes encoding molecules involved in responses to stimuli and muscle-related development were abundantly expressed in the oral sucker. Compared with other species, genes encoding molecules that facilitate the recognition and transport of cholesterol were observed in high copy numbers in the genome and were highly expressed in the oral sucker. Genes encoding transporters for fatty acids, glucose, amino acids and oxygen were also highly expressed, along with other molecules involved in metabolizing these substrates. All genes involved in energy metabolism pathways, including the ß-oxidation of fatty acids, the citrate cycle, oxidative phosphorylation, and fumarate reduction, were expressed in the adults. Finally, we also provide valuable insights into the mechanism underlying the process of pathogenesis by characterizing the secretome of C. sinensis. The characterization and elaborate analysis of the upgraded genome and the tissue transcriptomes not only form a detailed and fundamental C. sinensis resource but also provide novel insights into the physiology and pathogenesis of C. sinensis. We anticipate that this work will aid the development of innovative strategies for the prevention and control of clonorchiasis.


Assuntos
Transformação Celular Neoplásica , Clonorchis sinensis/genética , Fígado/parasitologia , Animais , Clonorquíase/parasitologia , Clonorchis sinensis/metabolismo , Clonorchis sinensis/patogenicidade , Análise por Conglomerados , Metabolismo Energético , Evolução Molecular , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Genoma de Protozoário , Humanos , Redes e Vias Metabólicas , Anotação de Sequência Molecular , Fases de Leitura Aberta , Especificidade de Órgãos/genética , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Transcriptoma
16.
Exp Parasitol ; 133(1): 62-9, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23149166

RESUMO

Ubiquitin is a functionally important protein expressed in eukaryotic cells usually encoded by multigenic families containing two types of genes, ubiquitin extension genes and polyubiquitin genes. One independent monomeric locus and two independent polyubiquitin loci were firstly identified from the genome of carcinogenic liver fluke, Clonorchis sinensis (C. sinensis). The nucleotide and amino acid sequence of C. sinensis polyubiquitin, especially polyubiquitin with five tandem ubiquitin repeats (CsPUB5), were analyzed. We obtained recombinant CsPUB5 (rCsPUB5) and anti-rCsPUB5 IgG. The ubiquitin transcripts in life cycle of C. sinensis were investigated. In addition, we found that ubiquitin or ubiquitination was ubiquitous in adult worm of C. sinensis and significantly observed in the content of biliary tract and intrahepatic biliary epithelium of liver from C. sinensis infected rat. We confirmed that rCsPUB5 could bind to human intrahepatic biliary epithelial cell by immunofluorescence in vitro. It was considered that ubiquitin family constitutively expressed in C. sinensis for variety of cellular processes and might be implicated in the genesis and progression of cholangiocarcinoma induced by the infection of C. sinensis.


Assuntos
Clonorchis sinensis/química , Ubiquitina/química , Ubiquitina/isolamento & purificação , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Gatos , Clonorchis sinensis/crescimento & desenvolvimento , DNA Complementar/química , Imunofluorescência , Humanos , Imunoglobulina G/imunologia , Estágios do Ciclo de Vida/genética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Ratos , Ratos Sprague-Dawley , Alinhamento de Sequência , Ubiquitina/genética , Ubiquitina/imunologia , Ubiquitinação
17.
Parasitol Res ; 111(3): 1143-50, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22752695

RESUMO

Clonorchiasis, caused by Clonorchis sinensis infection, has been an important public health problem in China. More messages about biology and pathogenicity of C. sinensis will be better for development of new strategies for clonorchiasis control. In the current study, using data from the published genomic and cDNA sequences for C. sinensis, we identify a gene encoding a myophilin-like protein (Cs myophilin-like). The cDNA sequence is 573 bp in length which encoded a protein of 190 amino acids. DNA sequence of Cs myophilin-like consists of four extrons and three introns. By mapping the draft genome of C. sinensis, single copy of myophilin-like gene is annotated. Bioinformatic analysis shows Cs myophilin-like comprises a calponin homology domain and three actin binding surfaces. It expresses at the life stage of adult worm and metacercaria but not in eggs. It intensively distributes in muscular locations such as oral sucker and pharynx of adult worm and sucker of metacercaria. The transcript level of Cs myophilin-like in oral sucker and pharynx is significantly higher than that in the rest of adult worm. Moreover, recombinant Cs myophilin-like is obtained and of antigenicity and immunoreactivity. It can be probed by sera from patients infected with trematode or cestode. Our data suggests that Cs myophilin-like may be involved in the host tissue invasion besides regulating the contraction of smooth muscle and movement of worm body. Taken together, the stable and soluble expression in Escherichia coli and immunogenicity of recombinant Cs myophilin-like proteins suggest that Cs myophilin-like may be a potential candidate for vaccine and drug target for clonorchiasis.


Assuntos
Antígenos de Helmintos/metabolismo , Clonorquíase/parasitologia , Clonorchis sinensis/metabolismo , Proteínas de Helminto/metabolismo , Estágios do Ciclo de Vida/fisiologia , Sequência de Aminoácidos , Animais , Clonorchis sinensis/genética , Proteínas de Helminto/genética , Humanos , Dados de Sequência Molecular
18.
Parasitol Res ; 111(3): 983-9, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22538482

RESUMO

Protein disulfide isomerase (PDI) is an essential catalyst of the endoplasmic reticulum with folding and chaperone activities in different biological systems. Here, PDI of Clonorchis sinensis (CsPDI) was isolated from the cDNA library of adult C. sinensis. The open reading frame contains 1,317 bp encoding 438 amino acids and shares 53 %, 49 %, and 43 % identity with PDI from Bos taurus, Homo sapiens, and Schistosoma mansoni, respectively. Two catalytic thioredoxin motifs CxxC were found in this sequence, which were characteristic domains of thioredoxin superfamily. The CsPDI protein was expressed and purified from Escherichia coli BL21 (DE3). According to western blotting analysis, the recombinant CsPDI could be recognized by anti-CsPDI rat serum, anti-excretory/secretory products rat serum, and serum of rat infected with C. sinensis, respectively. Quantitative real-time polymerase chain reaction showed that transcription level of CsPDI in the metacercaria stage was six and four times higher than that in the adult worm and egg stage, respectively. Immunolocalization analysis showed CsPDI could be detected in the intestine, vitellarium, and intrauterine eggs of adult worm, as well as in the cyst wall and vitellarium of metacercaria. In addition, the strong fluorescence signal was observed both on the wall of bile duct and in the lumen of liver tissue of C. sinensis-infected cat. Those results demonstrated that CsPDI was a component of C. sinensis excretory-secretory products. The present study will enhance our understanding of biological functions of CsPDI and pave the way for further studies on host-parasite interaction during C. sinensis infection.


Assuntos
Clonorchis sinensis/enzimologia , Regulação Enzimológica da Expressão Gênica/fisiologia , Sequência de Aminoácidos , Animais , Anticorpos Anti-Helmínticos , Ductos Biliares/patologia , Western Blotting , Doenças do Gato/parasitologia , Gatos , Clonagem Molecular , Clonorquíase/parasitologia , Clonorquíase/patologia , Clonorquíase/veterinária , Clonorchis sinensis/genética , Fígado/patologia , Hepatopatias/parasitologia , Hepatopatias/patologia , Hepatopatias/veterinária , Dados de Sequência Molecular , Ratos , Reação em Cadeia da Polimerase em Tempo Real
19.
Beilstein J Org Chem ; 8: 170-6, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22423284

RESUMO

Five carbamate derivatives, obtucarbamates C and D (1, 2), dimethyl ((carbonylbis(azanediyl))bis(2-methyl-5,1-phenylene))dicarbamate (3), obtucarbamates A and B (4, 5), and four aromadendrane-type sesquiterpenoids, (+)-4ß-N-methenetauryl-10ß-methoxy-1ß,5α,6ß,7ß-aromadendrane (6), (-)-4ß-N-methenetauryl-10ß-methoxy-1ß,5ß,6α,7α-aromadendrane (7), (-)-4α,10ß-aromadendranediol (8), (+)-4ß,10ß-aromadendranediol (9) were obtained from the South China Sea gorgonian coral Melitodes squamata Nutting. Compounds 1, 2, 6, and 7 were new, and their structures were established by spectroscopic analyses. Compounds 6 and 7 contained a taurine group that was rarely found in marine natural compounds, and 7 showed moderate antibacterial activity. The possible biosynthesis routes of 1-5 were conjectured.

20.
Mol Cell Biochem ; 363(1-2): 367-76, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22189506

RESUMO

Clonorchis sinensis fatty acid-binding protein (CsFABP) belongs to a multigene family of lipid-binding proteins and is considered to be a promising vaccine candidate for human clonorchiasis. In this study, binding characteristics of CsFABP have been examined for the first time. The recombinant CsFABP (rCsFABP) was found to bind 11-(dansylamino) undecanoic acid (DAUDA), causing a blue shift in the fluorescence emission from 543 to 531 nm with an excitation wavelength of 345 nm and a substantial increase in fluorescence intensity. Fluorimetric titration of rCsFABP with DAUDA exhibited an apparent dissociation constant (K (d)) of 1.58 ± 0.14 µM. In the competitive experiment, the rCsFABP efficiently bound saturated C(10)-C(18) fatty acids and unsaturated fatty acids (oleic acid and linoleic acid), and the latter presented the higher affinity. Furthermore, quantitative RT-PCR and western blotting analysis revealed that CsFABP mRNA and protein were differentially expressed throughout the developmental cycle stages of the parasite, which occur in the definitive host (metacercariae, adult worms, and eggs). In addition, immunolocalization assay showed that CsFABP was localized on the vitelline gland, tegument, intestine, seminal vesicle, eggs in uterus, ovary, and testicle of C. sinensis adult worm, as well as on the vitelline gland of metacercaria. Intriguingly, the surface tissue of the bile duct where C. sinensis resided in the infected Sprague-Dawley rat was also strongly labeled, implying that CsFABP may possibly mediate direct interactions with host cells as a component of excretory/secretory products.


Assuntos
Clonorchis sinensis/metabolismo , Proteínas de Ligação a Ácido Graxo/metabolismo , Proteínas de Helminto/metabolismo , Animais , Ductos Biliares/metabolismo , Ductos Biliares/parasitologia , Ligação Competitiva , Western Blotting , Clonorquíase/metabolismo , Clonorquíase/parasitologia , Clonorchis sinensis/genética , Clonorchis sinensis/crescimento & desenvolvimento , Compostos de Dansil/metabolismo , Modelos Animais de Doenças , Proteínas de Ligação a Ácido Graxo/química , Proteínas de Ligação a Ácido Graxo/genética , Ácidos Graxos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Helminto/genética , Modelos Moleculares , Reação em Cadeia da Polimerase , Conformação Proteica , RNA de Helmintos/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/metabolismo , Espectrometria de Fluorescência
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