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1.
Viral Immunol ; 2019 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-31381497

RESUMO

The human papillomavirus (HPV) vaccine has not been widely used in developing countries because of its high cost and multiple subtype restrictions. The present study aimed to develop an economical, convenient, and effective vaccine to produce neutralizing antibodies. Using late protein 1 (L1) from the HPV16 subtype as the target antigen (HPV16L1) and Pichia pastoris as the antigen release system, integrated P. pastoris expressing HPV16L1 (named yeast-HPV16L1) was prepared and vaccinated directly into mice by subcutaneous multipoint injection. After immunization was performed thrice, high titers (greater than 1:40,960) of specific anti-HPV16L1 antibodies were obtained in immune serum and were observed to continuously rise over time. The indirect hemagglutination test and indirect hemagglutination inhibition test were used to detect neutralizing antibody activity in vitro, and the results demonstrated the hemagglutination ability of the immune serum and the reduction in or loss of the hemagglutination ability if preneutralized antigen was added to the immune serum. The protection conferred by immune serum to tumor-bearing mice at the early stages was confirmed, but the neutralizing activity disappeared when the tumor reached a size of 1 mm3. The neutralization activity of the immune serum was confirmed both in vitro and in vivo.

2.
Artigo em Inglês | MEDLINE | ID: mdl-31329547

RESUMO

Person Re-identification (re-id) aims to match people across non-overlapping camera views in a public space. It is a challenging problem because many people captured in surveillance videos often wear similar clothes. Consequently, the differences in their appearance are typically subtle and only detectable at the right locations and scales. In this paper, a deep re-id network is proposed consisting of two novel components: a multi-scale deep learning layer and a leader-based attention learning layer. With these components, our model is able to learn deep discriminative feature representations at different scales and automatically determine the optimal weightings for each scale when combining them. The importance of different spatial locations for extracting discriminative features is also learned explicitly via our leader-based attention module. Extensive experiments are carried out to demonstrate that the proposed model outperforms the state-of-the-art on a number of benchmarks, and has a better generalization ability under a domain generalization setting.

3.
BMC Infect Dis ; 19(1): 512, 2019 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-31182037

RESUMO

BACKGROUND: The methods routinely used to detect trichomonads in the lungs are not sensitive enough, and an effective method is urgently needed. METHOD: Primers were first designed to match the conserved area of the 18S rRNA gene of trichomonads. Then, nested PCR was carried out to detect trichomonads in bronchoalveolar lavage fluid (BALF). Finally, all positive specimens were subjected to DNA sequencing and phylogenetic analysis. RESULTS: Among 115 bronchoalveolar lavage fluid samples, ten samples tested positive in nested PCR (10/115), while no samples were positive in wet mount microscopy (0/115) (P < 0.01). Among the ten positive specimens, two were identified as Tetratrichomonas spp. and the other eight as Trichomonas tenax in phylogenetic analysis. CONCLUSIONS: Nested PCR is an effective way to detect trichomonads in bronchoalveolar lavage fluid.


Assuntos
Líquido da Lavagem Broncoalveolar/microbiologia , Reação em Cadeia da Polimerase/métodos , Trichomonas/genética , DNA/química , DNA/metabolismo , Humanos , Filogenia , Análise de Sequência de DNA , Trichomonas/classificação , Trichomonas/isolamento & purificação , Tricomoníase/diagnóstico , Tricomoníase/microbiologia
4.
Artigo em Inglês | MEDLINE | ID: mdl-31199251

RESUMO

Despite significant progress in object categorization, in recent years, a number of important challenges remain; mainly, ability to learn from limited labeled data and ability to recognize object classes within large, potentially open, set of labels. Zero-shot learning is one way of addressing these challenges, but it has only been shown to work with limited sized class vocabularies and typically requires separation between supervised and unsupervised classes, allowing former to inform the latter but not vice versa. We propose the notion of vocabulary-informed learning to alleviate the above mentioned challenges and address problems of supervised, zero-shot, generalized zero-shot and open set recognition using a unified framework. Specifically, we propose a weighted maximum margin framework for semantic manifold-based recognition that incorporates distance constraints from (both supervised and unsupervised) vocabulary atoms. Distance constraints ensure that labeled samples are projected closer to their correct prototypes, in the embedding space, than to others. We illustrate that resulting model shows improvements in supervised, zero-shot, generalized zero-shot, and large open set recognition, with up to 310K class vocabulary on AwA and ImageNet datasets.

5.
Artigo em Inglês | MEDLINE | ID: mdl-31199252

RESUMO

We propose Deeply Supervised Object Detectors (DSOD), an object detection framework that can be trained from scratch. Recent advances in object detection heavily depend on the off-the-shelf models pre-trained on large-scale classification datasets like ImageNet and OpenImage. However, one problem is that adopting pre-trained models from classification to detection task may incur learning bias due to the different objective function and diverse distributions of object categories. Techniques like fine-tuning on detection task could alleviate this issue to some extent but are still not fundamental. Furthermore, transferring these pre-trained models cross discrepant domains will be more difficult (e.g., from RGB to depth images). Thus, a better solution to handle these critical problems is to train object detectors from scratch, which motivates our proposed method. In DSOD, we contribute a set of design principles for learning object detectors from scratch. One of the key principles is the deep supervision, enabled by layer-wise dense connections in both backbone networks and prediction layers, plays a critical role in learning good detectors from scratch. We evaluate our method on PASCAL VOC 2007, 2012 and COCO datasets. DSOD achieves consistently better results than the state-of-the-art methods with much more compact models.

6.
Artigo em Inglês | MEDLINE | ID: mdl-30969924

RESUMO

The ability to quickly recognize and learn new visual concepts from limited samples enables humans to quickly adapt to new tasks and environments. This ability is enabled by semantic association of novel concepts with those that have already been learned and stored in memory. Computers can start to ascertain similar abilities by utilizing a semantic concept space. A concept space is a high-dimensional semantic space in which similar abstract concepts appear close and dissimilar ones far apart. In this paper, we propose a novel approach to one-shot learning that builds on this core idea. Our approach learns to map a novel sample instance to a concept, relates that concept to the existing ones in the concept space and, using these relationships, generates new instances, by interpolating among the concepts, to help learning. Instead of synthesizing new image instance, we propose to directly synthesize instance features by leveraging semantics using a novel auto-encoder network we call dual TriNet. The encoder part of the TriNet learns to map multi-layer visual features from CNN to a semantic vector. In semantic space, we search for related concepts, which are then projected back into the image feature spaces by the decoder portion of the TriNet. Two strategies in the semantic space are explored. Notably, this seemingly simple strategy results in complex augmented feature distributions in the image feature space, leading to substantially better performance. The codes and models are released in the github: https://github.com/tankche1/ Semantic-Feature-Augmentation-in-Few-shot-Learning.

7.
J Transl Med ; 16(1): 370, 2018 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-30577810

RESUMO

BACKGROUND: Systemic lupus erythematosus (SLE) is a multisystemic autoimmune disease with various clinical manifestations. MicroRNAs (miRNAs) and immunometabolism are recognized as key elements in SLE pathogenesis; however, the relationship between miRNAs in peripheral blood mononuclear cells (PBMCs) and metabolism in SLE remains unclear. METHODS: We detected PBMC miRNA and mRNA profiles from 3 pooled SLE patients and 3 healthy controls (HCs) using next-generation sequencing, predicted miRNA targets in dysregulated mRNAs, predicted functions and interactions of differentially expressed genes using bioinformatics analysis, validated candidate miRNAs using qRT-PCR, and investigated the association between the expression of candidate miRNAs and SLE clinical characteristics. Moreover, we validated the direct and transcriptional regulatory effect of NovelmiRNA-25 on adenosine monophosphate deaminase 2 (AMPD2) using a dual-luciferase reporter assay and western blot and confirmed AMPD2 mRNA and protein expression in PBMCs using qRT-PCR and western blot, respectively. RESULTS: Multilayer integrative analysis of microRNA and mRNA regulation showed that 10 miRNAs were down-regulated and 19 miRNAs were up-regulated in SLE patient PBMCs compared with HCs. Bioinformatics analysis of regulatory networks between miRNAs and mRNAs showed that 19 miRNAs were related to metabolic processes. Two candidate miRNAs, NovelmiRNA-25 and miR-1273h-5p, which were significantly increased in the PBMCs of SLE patients (P < 0.05), represented diagnostic biomarkers with sensitivities of 94.74% and 89.47%, respectively (area under the curve = 0.574 and 0.788, respectively). NovelmiRNA-25 expression in PBMCs was associated with disease activity in SLE patients, in both active and stable groups (P < 0.05). NovelmiRNA-25 overexpression downregulated AMPD2 expression in HEK293T cells through direct targeting of the AMPD2 3'UTR (P < 0.01), while inhibition of NovelmiRNA-25 activity led to increased AMPD2 expression (P < 0.01). NovelmiRNA-25 overexpression also downregulated AMPD2 protein expression in HEK293T cells; AMPD2 protein expression in SLE patient PBMCs was decreased. Our results show that differentially expressed miRNAs play an important role in SLE. CONCLUSIONS: Our data demonstrate a novel mechanism in SLE development that involves the targeting of AMPD2 expression by NovelmiRNA-25. miRNAs may serve as novel biomarkers for the diagnosis and evaluation of disease activity of SLE and represent potential therapeutic targets for this disease.


Assuntos
AMP Desaminase/sangue , Leucócitos Mononucleares/metabolismo , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/genética , MicroRNAs/metabolismo , Sequência de Bases , Biomarcadores/sangue , Estudos de Casos e Controles , Ontologia Genética , Redes Reguladoras de Genes , Células HEK293 , Humanos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
8.
Oncol Lett ; 16(4): 4372-4378, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30214572

RESUMO

Epstein-Barr virus (EBV) is widespread and is associated with nasopharyngeal carcinoma (NPC). Serological detection of EBV is commonly used for screening, diagnosis and epidemiological surveys of NPC. In the present study, a novel B cell multi-epitope peptide fusion protein (EBV-LMP2-3B), which is composed of three B cell linear epitopes (RIEDPPFNSLL, TLNLT and KSLSSTEFIPN) of EBV latent membrane protein 2 (LMP2), was expressed in a prokaryotic expression system and purified using Ni2+-nitrilotriacetate-Sepharose. The immunogenicity and binding specificity of EBV-LMP2-3B were evaluated on the basis of antibody responses in immunized BALB/c mice, western blotting and indirect immunofluorescence assay. Evaluation of EBV-LMP2-3B as a serological diagnostic reagent was performed using an indirect ELISA in 198 patients with NPC and 102 healthy adults. These results revealed that EBV-LMP2-3B was able to eliminate the high-titer serum antibody response in BALB/c mice. Western blot analysis and indirect immunofluorescence assay confirmed that the mouse immune sera recognized the native LMP2. Compared with healthy adults, patients with NPC demonstrated significantly greater reactivity to EBV-LMP2-3B (P<0.05). Furthermore, it was possible to effectively detect specific IgG in sera from patients with NPC, with a sensitivity of 91.91% and specificity of 93.14%, representing an improvement over the traditional viral capsid antigen-IgA-based detection method with 59.59% sensitivity and 75.49% specificity. In conclusion, the EBV-LMP2-3B protein may be used as a serological diagnostic reagent to screen for and diagnose NPC.

9.
Front Immunol ; 9: 1473, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30008716

RESUMO

Our understanding of circulating microRNAs (miRNAs) related to systemic lupus erythematosus (SLE) remains very limited. In this study, we screened SLE-specific miRNAs in plasma from 42 B cell-related miRNAs by using miRNA PCR Array. The selected miRNAs were first confirmed in plasma samples from 50 SLE patients, 16 rheumatoid arthritis (RA) patients, and 20 healthy donors using qRT-PCR. We then investigated the relationship between expressions of the selected miRNAs and SLE clinical indicators. As a result, 14 miRNAs (miR-103, miR-150, miR-20a, miR-223, miR-27a, miR-15b, miR-16, miR-181a, miR-19b, miR-22, miR-23a, miR-25, miR-92a, and miR-93) were significantly decreased in the plasma of SLE patients compared with healthy controls (P < 0.05) and could act as the diagnostic signature to distinguish SLE patients from healthy donors. Six miRNAs (miR-92a, miR-27a, miR-19b, miR-23a, miR-223, and miR-16) expressed in plasma were significantly lower in SLE patients than in RA patients (P < 0.05), revealing the potentially diagnostic signature to distinguish SLE patients from RA patients. Furthermore, the downregulated expression of miR-19b, miR-25, miR-93, and miR-15b was associated with SLE disease activity (P < 0.05) while miR-15b and miR-22 expressions were significantly lower in SLE patients with low estimate glomerular filtration rate (eGFR < 60 ml/min/1.73 m2) (P < 0.05). The diagnostic potential of miR-15b for SLE disease activity and lupus nephritis (LN) with low eGFR was validated on an independent validation set with 69 SLE patients and a cross-validation set with 80 SLE patients. In summary, the signature of circulating miRNAs will provide novel biomarkers for the diagnosis of SLE and evaluation of disease activity and LN.

10.
Artigo em Inglês | MEDLINE | ID: mdl-29938318

RESUMO

High-risk human papillomavirus (HPV16 and HPV18) are now widely recognized as responsible for cervical cancer, which remains to be the most common gynecologic malignancy in women worldwide. It is well known that viral oncoproteins E6/E7 play key roles in HPV-associated cervical carcinogenesis. Thus, in vivo detection of the two oncoproteins may provide important diagnostic information influencing patient management. More recently, affibody molecules have been demonstrated to be a promising candidate for development as molecular imaging probes. Based on the two monomeric affibody molecules (ZHPV16E7 and ZHPV18E7) generated in our laboratory, here, we used a peptide linker (Gly4Ser)3 to link ZHPV16E7 and ZHPV18E7 to develop a novel heterodimeric affibody ZHPV16E7-(Gly4Ser)3-ZHPV18E7. Both biosensor and immunofluorescence assays have proved that the heterodimeric affibody molecule targeted simultaneously HPV16 and HPV18E7 proteins by binding to the viral oncoproteins. In vivo tumor-imaging experiments using the Dylight755-labeled heterodimeric affibody revealed that strongly high-contrast tumor retention of the heterodimers occurred in both HPV16- and HPV18-derived tumors of nude mice 0.5 h post-injection. The accumulation of Dylight755-labeled heterodimers in tumors was achieved over 48 h. Therefore, we believe that this novel heterodimeric affibody molecule has great potential utility in molecular imaging in vivo and diagnosis of HPV-associated cervical cancers.

11.
Diabetes Metab Syndr Obes ; 11: 277-288, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29942142

RESUMO

Background: The neutrophil-lymphocyte ratio (NLR) has been considered as an inflammatory marker in various disorders, but it is not clear whether the NLR is also elevated with hidden diabetes (HD), which is normal in fasting blood glucose (FBG) but abnormal in the oral glucose tolerance test (OGTT). Materials and methods: An HD animal model for 27 days and an animal model with oscillating glucose (OG) for 7 days were applied on adult female Sprague-Dawley rats. OGTT, leukogram analysis, histology, and immunohistochemistry were carried out. Results: In HD rats, the percentage of neutrophils increased but the percentage of lymphocytes decreased; hence, the NLR rose relative to sham. This may be a result of the OG levels often experienced by diabetic subjects, as normal rats given OG (6 g/kg/6 h) for 7 days had significantly reduced lymphocyte numbers and increased NLR compared with the values before and 1 h after oral glucose administration during OGTT. Glucose-induced disarrangement of partitions of circulating immune cells and NLR was involved in the increase in oxidative stress, as these changes were totally blocked by the antioxidant glutathione (GSH). GSH (50 mg/kg/6 h) totally blocked the glucose-induced alterations in lymphocyte and NLR values. Conclusion: HD associated with elevation of NLR values may be partly attributed to a homeostasis disorder of the innate inflammatory state, caused by oscillating hyperglycemia. Acute high glucose administration produced a significant decrease in lymphocyte number. OG administration potentiated this effect and increased the NLR value, which was blocked by GSH, suggesting that reactive oxygen species play a critical role in maintaining lymphocyte numbers.

12.
Front Immunol ; 9: 187, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29497417

RESUMO

This study was aimed to evaluate the role of B-cell epitopes of Epstein-Barr virus (EBV) Early antigen protein D (EA), envelope glycoprotein GP340/membrane antigen (MA), latent membrane protein (LMP)-1, and LMP-2A in systemic lupus erythematosus (SLE). B-cell epitopes were predicted by analyzing secondary structure, transmembrane domains, surface properties, and homological comparison. 60 female mice were randomized equally into 12 groups: 1-10 groups were immunized by epitope peptides (EPs) 1-10, respectively, while 11 and 12 groups were PBS and Keyhole limpet hemocyanin (KLH) control groups. Immunoglobulin G (IgG) and autoantibody to nuclear antigen (ANA) concentrations in mice serum were determined at week 8. Indirect levels of EP1-10 were further detected by enzyme-linked immuno sorbent assay (ELISA) in 119 SLE patients and 64 age- and gender-matched health controls (HCs). 10 probable EBV EA, MA, LMP-1, and LMP-2A B-cell epitopes related to SLE self-antigens were predicted and corresponding EP1-10 were synthesized. IgG concentrations at week 8 were increased in EP1-10 and KLH groups compared with PBS group in mice; while ANA levels were elevated in only EP1-4, EP6-7, and EP10 groups compared to KLH group by ELISA, and ANA-positive rates were increased in only EP1, EP2, EP4, EP6, and EP10 groups by indirect immunofluorescence assay. EP1-4, EP6, and EP10 indirect levels were increased in SLE patients than HCs, while EP1, EP3, EP6, and EP9 were correlated with SLE disease activity index score. In conclusion, EBV EA, MA, LMP-1, and LMP-2A B-cell EPs increased SLE-related autoantibodies in mice, and their indirect levels might be served as potential biomarkers for SLE diagnosis and disease severity.

13.
Cell Death Dis ; 9(2): 104, 2018 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-29367719

RESUMO

It has been hypothesized that human cytomegalovirus (HCMV) infection, especially in monocyte and CD34 (+) myeloid cells, acts as a important regulator of immune system to promote inflammation in multiple autoimmune diseases. The aim of this study was to elucidate the HCMV gene expression profiles in the peripheral blood mononuclear cells (PBMCs) of SLE patients and demonstrate the effect and mechanism of viral gene associated with SLE in mono-macrophages functions. Using two RNA-Seq techniques in combination with RT-PCR, 11 viral genes mainly associated with latent HCMV infection were identified in the PBMCs of SLE patients. Among these viral genes, US31 with previously unknown function was highly expressed in the PBMCs of SLE patients compared to healthy controls. Analysis of function indicated that US31 expression could induce inflammation in monocyte and macrophage and stimulate macrophage differentiation toward an M1 macrophage phenotype. Screening via protein chips in combination with bioinformatic analysis and consequent detection of mono-macrophages function indicates that the direct interaction between US31 and NF-κB2 contributed the NF-kB2 activation. Consequent analysis indicated US31 directly interacted with NF-κB2, contribute to the polyubiquitination of the phosphorylated p100 and consequent activation of NF-κB2. Taken together, our data uncovered a previously unknown role of the HCMV protein US31 in inducing NF-κB-mediated mono-macrophage inflammation in the pathogenesis and development of SLE. Our findings provide a foundation for the continued investigation of novel therapeutic targets for SLE patients.

14.
IEEE Trans Pattern Anal Mach Intell ; 40(2): 352-364, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28221992

RESUMO

In this paper, we study the challenging problem of categorizing videos according to high-level semantics such as the existence of a particular human action or a complex event. Although extensive efforts have been devoted in recent years, most existing works combined multiple video features using simple fusion strategies and neglected the utilization of inter-class semantic relationships. This paper proposes a novel unified framework that jointly exploits the feature relationships and the class relationships for improved categorization performance. Specifically, these two types of relationships are estimated and utilized by imposing regularizations in the learning process of a deep neural network (DNN). Through arming the DNN with better capability of harnessing both the feature and the class relationships, the proposed regularized DNN (rDNN) is more suitable for modeling video semantics. We show that rDNN produces better performance over several state-of-the-art approaches. Competitive results are reported on the well-known Hollywood2 and Columbia Consumer Video benchmarks. In addition, to stimulate future research on large scale video categorization, we collect and release a new benchmark dataset, called FCVID, which contains 91,223 Internet videos and 239 manually annotated categories.

15.
PLoS Pathog ; 13(11): e1006715, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29176795

RESUMO

Mouse papillomavirus type 1 (MmuPV1) provides, for the first time, the opportunity to study infection and pathogenesis of papillomaviruses in the context of laboratory mice. In this report, we define the transcriptome of MmuPV1 genome present in papillomas arising in experimentally infected mice using a combination of RNA-seq, PacBio Iso-seq, 5' RACE, 3' RACE, primer-walking RT-PCR, RNase protection, Northern blot and in situ hybridization analyses. We demonstrate that the MmuPV1 genome is transcribed unidirectionally from five major promoters (P) or transcription start sites (TSS) and polyadenylates its transcripts at two major polyadenylation (pA) sites. We designate the P7503, P360 and P859 as "early" promoters because they give rise to transcripts mostly utilizing the polyadenylation signal at nt 3844 and therefore can only encode early genes, and P7107 and P533 as "late" promoters because they give rise to transcripts utilizing polyadenylation signals at either nt 3844 or nt 7047, the latter being able to encode late, capsid proteins. MmuPV1 genome contains five splice donor sites and three acceptor sites that produce thirty-six RNA isoforms deduced to express seven predicted early gene products (E6, E7, E1, E1^M1, E1^M2, E2 and E8^E2) and three predicted late gene products (E1^E4, L2 and L1). The majority of the viral early transcripts are spliced once from nt 757 to 3139, while viral late transcripts, which are predicted to encode L1, are spliced twice, first from nt 7243 to either nt 3139 (P7107) or nt 757 to 3139 (P533) and second from nt 3431 to nt 5372. Thirteen of these viral transcripts were detectable by Northern blot analysis, with the P533-derived late E1^E4 transcripts being the most abundant. The late transcripts could be detected in highly differentiated keratinocytes of MmuPV1-infected tissues as early as ten days after MmuPV1 inoculation and correlated with detection of L1 protein and viral DNA amplification. In mature warts, detection of L1 was also found in more poorly differentiated cells, as previously reported. Subclinical infections were also observed. The comprehensive transcription map of MmuPV1 generated in this study provides further evidence that MmuPV1 is similar to high-risk cutaneous beta human papillomaviruses. The knowledge revealed will facilitate the use of MmuPV1 as an animal virus model for understanding of human papillomavirus gene expression, pathogenesis and immunology.


Assuntos
Papillomaviridae/genética , Infecções por Papillomavirus/virologia , Doenças dos Roedores/virologia , Proteínas Virais/genética , Verrugas/veterinária , Animais , Feminino , Genoma Viral , Camundongos , Camundongos Endogâmicos BALB C , Papillomaviridae/metabolismo , RNA Viral/genética , RNA Viral/metabolismo , Transcriptoma , Proteínas Virais/metabolismo , Verrugas/virologia
16.
Oncotarget ; 8(38): 62914-62926, 2017 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-28968959

RESUMO

Aberrantly expressed microRNAs contribute to the initiation and progression of human cancer. MiRNA-187 has been reported in nasopharyngeal, renal, pancreatic, prostate, and esophageal cancer, and acts as a tumor suppressor or oncogene. However, the underlying function of miRNA-187 in cervical cancer remains largely unexplored. In the present study, we demonstrated significantly miRNA-187 down-regulation in cervical cancer tissues and cell lines compared to their normal counterparts. Kaplan-Meier analysis revealed that decreased miRNA-187 was closely associated with shorter overall survival and relapse-free survival. Gain- and loss-of-function studies showed that miRNA-187 suppressed cervical cancer cell proliferation, migration, and invasion, and promoted cervical cancer cell apoptosis. Furthermore, luciferase reporter assay determined that human papillomavirus 16 E6 was a direct functional target of miRNA-187. Taken together, our findings indicate the essential role of miRNA-187 in suppressing cervical cancer progression and indicate a novel link between miRNA-187 and human papillomavirus 16 E6 in cervical cancer.

17.
Cancer Med ; 6(9): 2063-2075, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28812328

RESUMO

In addition to regulating apoptosis via its interaction with the death domain of Fas receptor, death domain associated protein 6 (Daxx) is also known to be involved in transcriptional regulation, suggesting that the function of Daxx depends on its subcellular localization. In this study, we aimed to explore Daxx subcellular localization in gastric cancer (GC) cells and correlate the findings with clinical data in GC patients. Seventy pairs of tissue samples (GC and adjacent normal tissue) were analyzed immunohistochemically for Daxx expression and localization (nuclear and cytoplasmic). The Daxx Nuclear/Cytoplasmic ratio (Daxx NCR) values in tissue microarray data with 522 tumor samples were further analyzed. The defined Prior cohort (n = 277, treatment between 2006 and 2009) and Recent cohort (n = 245, treatment between 2010 and 2011) were then used to examine the relationship between Daxx NCR and clinical data. The Daxx NCR was found to be clinically informative and significantly higher in GC tissue. Using Daxx NCR (risk ratio = 2.0), both the Prior and Recent cohorts were divided into high- and low-risk groups. Relative to the low-risk group, the high-risk patients had a shorter disease free survival (DFS) and overall survival (OS) in both cohorts. Importantly, postoperative chemotherapy was found having differential effect on high- and low-risk patients. Such chemotherapy brought no survival benefit, (and could potentially be detrimental,) to high-risk patients after surgery. Daxx NCR could be used as a prognosis factor in GC patients, and may help select the appropriate population to benefit from chemotherapy after surgery.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Proteínas Nucleares/metabolismo , Neoplasias Gástricas/patologia , Análise Serial de Tecidos/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Estudos Retrospectivos , Neoplasias Gástricas/metabolismo , Análise de Sobrevida , Regulação para Cima , Adulto Jovem
18.
Protein Pept Lett ; 24(10): 947-954, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28641561

RESUMO

BACKGROUND: Melanoma-associated antigen-A3 (MAGE-A3) is a tumor specific antigen and a potential candidate for cancer immunotherapy. We had screened three immunodominant multiepitopes of MAGE-A3, and identified these multiepitope peptides had significantly higher reactivity to serum samples from gastric cancer patients. However, the immune responses of three multiepitope peptides carried by HBcAg in mice have not been investigated. OBJECTIVES: The main objective of this study was to analyze the humoral and cellular immune responses in mice induced by these three multiepitope vaccines of MAGE-A3. METHODS: Three multiepitopes of MAGE-A3 (MAGE-A3(EPI-1, or -2, or -3)) were respectively inserted at HBcAg major immunodominant region (HBcAg(MIR)) of the pET21a(+)/HBcAg(MIR) recombinant plasmid. These recombinant chimeras were identified by PCR, and transfected respectively into E. Coli Ressotta strain. The expression products of rHBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) were purified respectively by Ni2+ chelated affinity column, and then confirmed by SDS-PAGE and Western-blot analysis.Purified three rHBcAg(MIR)/MAGE-A3 multiepitopes were administrated respectively into BALB/c (H-2Kd) mice by intradermal injection. The production of rHBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) specific IgG in serum from immunized mice were measured by ELISA. Spleen cells from all immunized mice were harvested after one week of last immunization for lymphocyte proliferation assay and cytotoxic T-lymphocyte assay. RESULTS: PCR and Sequencing analysis showed the presence of the required gene fragment in pET21a(+)/ HBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) recombinant plasmid. Purified rHBcAg(MIR)/MAGE-A3(EPI-1, or -2, or -3) could be probed specifically by McAb of 6×his-tag. ELISA analysis indicated that serum from immunized mice with rHBcAg(MIR)/MAGE-A3(EPI-1, -2, or -3) proteins could be discerned specifically by complete MAGE-A3 protein, and high level of antibodies in immune serum were obtained, and all antibody titers could reach above 1:1600. The splenocytes from groups of rHBcAg(MIR)/MAGE-A3(EPI-1,-2, or -3), stimulated respectively with corresponding peptides showed the higher proliferative responses comparing with control groups of HBcAg(MIR) or PBS (p<0.05, respectively). Splenocytes from mice immunized with rHBcAg(MIR)/MAGE-A3 (EPI-1, or -2, or -3) could killed target cells effectively, and there were significant difference of CTL activities compared with control groups of HBcAg(MIR), or PBS (p<0.05, respectively) at any ratio of effector : target. CONCLUSION: Our results indicated MIR in HBcAg presenting platform could present MAGE-A3 multiepitopes efficiently and induced significant humoral or cellular immunity. The immune strategy based on multiepitopeimmunization could have potential for preventing or controlling MAGE-A3 associated malignant disease.

19.
Vaccine ; 35(23): 3096-3103, 2017 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-28456528

RESUMO

Chlamydia trachomatis (Ct), an obligate intracellular parasite, is the leading cause of bacterial sexually transmitted diseases worldwide. The best solution to control the spread of Ct is to develop safe and effective vaccines. However, an effective vaccine has not been developed due to some challenges such as selection of appropriate candidate antigens and an effective delivery system. In our previous study, we have developed a Ct vaccine that comprises a multi-epitope peptide of Ct major outer membrane protein (MOMP370-387) and Hepatitis B virus core antigen (HBcAg). The vaccine was evaluated in a murine model with chlamydial genital infection. The results indicated that Ct MOMP multi-epitope delivered by HBcAg could be an effective vaccine for the prevention of Ct. In this study, another two epitopes were selected from the MOMP protein and tandemly linked with MOMP370-387 to enhance the immunogenicity and the protective effect of the candidate vaccine. Our results revealed that both the immunogenicity and the protective effect of the tandem Ct MOMP multi-epitopes were much better than that of the single epitope. Therefore, vaccines based on the tandem Ct MOMP multi-epitopes could be more effective immune prophylactics to prevent Ct infection than the single epitope in murine model system.


Assuntos
Vacinas Bacterianas/imunologia , Infecções por Chlamydia/prevenção & controle , Chlamydia trachomatis/imunologia , Epitopos/imunologia , Porinas/imunologia , Animais , Anticorpos Antibacterianos/sangue , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/química , Infecções por Chlamydia/microbiologia , Chlamydia trachomatis/química , Modelos Animais de Doenças , Epitopos/química , Feminino , Imunogenicidade da Vacina , Camundongos , Camundongos Endogâmicos BALB C , Porinas/química
20.
Oncotarget ; 8(20): 33241-33251, 2017 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-28402260

RESUMO

Chlamydia trachomatis (Ct) is one of the most frequently encountered sexual infection all over the world, yielding tremendous reproductive problems (e.g. infertility and ectopic pregnancy) in the women. This work described the design of a plasmid vaccine that protect mice from Ct infection, and reduce productive tract damage by generating effective antibody and cytotoxic T cell immunity. The vaccine, s was composed of MOMP multi-epitope and HPV16L2 genes carried in pcDNA plasmid (i.e. pcDNA3.1/MOMP/HPV16L). In transfection, the vaccine expressed the chimeric genes (i.e. MOMP and HPV16L2), as demonstrated via western blot, RT-PCR and fluorescence imaging. In vitro, the vaccine transfected COS-7 cells and expressed the proteins corresponding to the genes carried in the vaccine. Through intramuscular immunization in BALB/c mice, the vaccine induced higher levels of anti-Ct IgG titer, anti-HPV16L2 IgG titer in serum and IgA titer in local mucosal secretions, compared to plasmid vaccines that carry only Ct MOMP multi-epitope or HPV16L2 chimeric component only. In mice intravaginally challenged with Ct, the vaccines pcDNA3.1/MOMP/HPV16L2 generated a higher level of genital protection compared to other vaccine formulations. Additionally, histochemical staining indicated that pcDNA3.1/MOMP/HPV16L2 eliminated mouse genital tract tissue pathologies induced by Ct infection. This work demonstrated that pcDNA/MOMP/HPV16L2 vaccine can protect against Ct infection by regulating antibody production, cytotoxic T cell killing functions and reducing pathological damage in mice genital tract. This work can potentially offer us a new vaccine platform against Ct infection.


Assuntos
Vacinas Bacterianas/imunologia , Proteínas do Capsídeo/imunologia , Infecções por Chlamydia/imunologia , Chlamydia trachomatis/imunologia , Proteínas Oncogênicas Virais/imunologia , Porinas/imunologia , Vacinas de DNA/imunologia , Animais , Anticorpos Antibacterianos/imunologia , Vacinas Bacterianas/administração & dosagem , Sequência de Bases , Células COS , Proteínas do Capsídeo/genética , Cercopithecus aethiops , Infecções por Chlamydia/prevenção & controle , Chlamydia trachomatis/genética , Epitopos/imunologia , Feminino , Expressão Gênica , Imunização , Camundongos , Proteínas Oncogênicas Virais/genética , Plasmídeos/administração & dosagem , Plasmídeos/genética , Plasmídeos/imunologia , Porinas/genética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Análise de Sequência de DNA , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismo , Transfecção , Vacinas de DNA/administração & dosagem
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