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1.
Med Sci Monit ; 27: e931590, 2021 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-33704261

RESUMO

The authors repeated experiments and found that the results shown in figure 2 were not reproducible. Reference: Shuang-li Zhang, Bao-lin Li, Wei Li, Ming Lu, Lin-ying Ni, Hui-li Ma, Qing-gang Meng. The Effects of Ludartin on Cell Proliferation, Cell Migration, Cell Cycle Arrest and Apoptosis Are Associated with Upregulation of p21WAF1 in Saos-2 Osteosarcoma Cells In Vitro. Med Sci Monit 2018; 24: LBR4926-4933. 10.12659/MSM.909193.

2.
J Orthop Surg Res ; 16(1): 21, 2021 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-33413520

RESUMO

BACKGROUND: There is now ample evidence suggesting that extracorporeal shock wave therapy (ESWT) can improve hip mobility and reduce pain in patients with osteonecrosis of the femoral head (ONFH). The ability of ESWT to cure bone marrow edema syndrome (BMES) in patients with ONFH, 12 weeks after the initial course of ESWT, needs to be verified further and more relevant clinical research-based evidence should be consolidated. This study aimed to evaluate the efficacy of ESWT for BMES caused by ONFH. METHODS: This retrospective cohort study included 67 patients with BMES caused by ONFH who were participating in a rehabilitation program as outpatients. Before and after ESWT, the area of femoral bone marrow edema was evaluated by magnetic resonance imaging (MRI), and the Harris score and Charnley score were evaluated as hip pain and function indicators. RESULTS: After ESWT, MRI revealed that the area of bone marrow edema decreased from 984.6 ± 433.2 mm2 to 189.7 ± 214.4 mm2 (P < 0.0001). The Harris score increased from 42.2 ± 9.1 to 77.7 ± 10.8 points (P < 0.0001). The Charnley score increased from 7.3 ± 1.4 to 12.0 ± 1.7 (P < 0.0001). ESWT was effective in treating BMES in 98.5% of the cases. CONCLUSIONS: This study demonstrated that ESWT can effectively treat BMES caused by ONFH and can aid in pain relief and functional recovery in patients with ONFH. Thus, ESWT should be included in the classic physical therapy regimen for patients with ONFH and BMES.

3.
IEEE Trans Cybern ; PP2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-33055048

RESUMO

Recently, the visual quality evaluation of screen content images (SCIs) has become an important and timely emerging research theme. This article presents an effective and novel blind quality evaluation metric for SCIs by using stacked autoencoders (SAE) based on pictorial and textual regions. Since the SCI consists of not only the pictorial area but also the textual area, the human visual system (HVS) is not equally sensitive to their different distortion types. First, the textual and pictorial regions can be obtained by dividing an input SCI via an SCI segmentation metric. Next, we extract quality-aware features from the textual region and pictorial region, respectively. Then, two different SAEs are trained via an unsupervised approach for quality-aware features that are extracted from these two regions. After the training procedure of the SAEs, the quality-aware features can evolve into more discriminative and meaningful features. Subsequently, the evolved features and their corresponding subjective scores are input into two regressors for training. Each regressor can obtain one output predictive score. Finally, the final perceptual quality score of a test SCI is computed by these two predicted scores via a weighted model. Experimental results on two public SCI-oriented databases have revealed that the proposed scheme can compare favorably with the existing blind image quality assessment metrics.

4.
IEEE Trans Cybern ; PP2020 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-32966229

RESUMO

While full automation of road vehicles remains a future goal, shared-control and semiautonomous driving--involving transitions of control between the human and the machine--are more feasible objectives in the near term. These alternative driving modes will benefit from new research toward novel steering control devices, more suitably where machine intelligence only partially controls the vehicle. In this article, it is proposed that when the human shares the control of a vehicle with an autonomous or semiautonomous system, a force control, or nondisplacement steering wheel (i.e., a steering wheel which does not rotate but detects the applied torque by the human driver) can be advantageous under certain schemes: tight rein or loose rein modes according to the H-metaphor. We support this proposition with the first experiments to the best of our knowledge, in which human participants drove in a simulated road scene with a force control steering wheel (FCSW). The experiments exhibited that humans can adapt promptly to force control steering and are able to control the vehicle smoothly. Different transfer functions are tested, which translate the applied torque at the FCSW to the steering angle at the wheels of the vehicle; it is shown that fractional order transfer functions increment steering stability and control accuracy when using a force control device. The transition of control experiments is also performed with both: a conventional and an FCSW. This prototypical steering system can be realized via steer-by-wire controls, which are already incorporated in commercially available vehicles.

5.
PeerJ ; 8: e8954, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32328351

RESUMO

Background: Osteosarcoma (OS) is the most common malignant bone tumor with high mortality in children and adolescents. REG γ is overexpressed and plays oncogenic roles in various types of human cancers. However, the expression and potential roles of REG γ in osteosarcoma are elusive. This study aims at exploring possible biological functions of REG γ in the pathogenesis of osteosarcoma and its underlying mechanism. Methods: Quantitativereverse transcription-polymerase chain reaction (qRT-PCR), western blotting andimmunohistochemistry (IHC)were performed to detect the expression levels of REG γ in OS tissues and cell lines. Then, the effects of REG γ expression on OS cell proliferation in vitro were analyzed by Cell Counting Kit-8 (CCK-8), ethylene deoxyuridine (EdU), colony formation, flow cytometry. The protein levels of apoptosis and cell-cycle related proteins were evaluated using western blotting. Results: In present study, we found for the first time that REG γ is overexpressed in osteosarcoma tissues and cell lines and knockdown of REG γ significantly inhibits cell proliferation and induces apoptosis and cell cycle arrest in osteosarcoma cells. Furthermore, we observed that p21, caspase-3 and cleaved caspase-3 are increased while the expression of cycinD1 and bcl-2 are decreased after REG γ depletion in osteosarcoma cells. In conclusion, REG γ may be involved in the proliferation of osteosarcoma and serve as a novel therapeutic target in patients with osteosarcoma.

6.
Am J Transl Res ; 12(2): 563-573, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32194904

RESUMO

BACKGROUND: Proteasome activator γ (REG γ) expression was found to be upregulated and to play critical roles in several cancers. However, the effect of REG γ on osteosarcoma (OS) remains unclear. The objective of the present study was to explore the clinical significance of REG γ and its function in regulating the progression of OS. METHODS: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting (WB) and immunohistochemistry (IHC) analyses were performed to detect the expression levels of REG γ in OS tissues and cell lines. Then, the effects of REG γ expression on OS cell behavior in vitro were analyzed by Cell Counting Kit-8 (CCK-8), ethylene deoxyuridine (EdU), colony formation, flow cytometry, wound healing and transwell assays. The protein and mRNA levels of components involved in the Wnt/ß-catenin pathway were evaluated using WB and qRT-PCR, respectively. RESULTS: We found that REG γ expression was significantly upregulated in both OS tissues and cell lines. Our in vitro assay results confirmed that knockdown of REG γ inhibited cell proliferation, migration, and invasion and induced apoptosis and cell cycle arrest in OS. Additionally, through WB and qRT-PCR analyses, we found that REG γ depletion markedly decreased the ß-catenin, cyclin D1 and c-myc expression levels and increased the GSK-3ß expression levels in OS cell lines. CONCLUSIONS: Our results revealed that REG γ plays an oncogenic role in OS by activating the Wnt/ß-catenin pathway, indicating that REG γ may be a promising therapeutic target for OS patients.

7.
IEEE Trans Cybern ; 50(1): 87-99, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30183651

RESUMO

An image is compressed or stretched during the multidevice displaying, which will have a very big impact on perception quality. In order to solve this problem, a variety of image retargeting methods have been proposed for the retargeting process. However, how to evaluate the results of different image retargeting is a very critical issue. In various application systems, the subjective evaluation method cannot be applied on a large scale. So we put this problem in the accurate objective-quality evaluation. Currently, most of the image retargeting quality assessment algorithms use simple regression methods as the last step to obtain the evaluation result, which are not corresponding with the perception simulation in the human vision system (HVS). In this paper, a deep quality evaluator for image retargeting based on the segmented stacked AutoEnCoder (SAE) is proposed. Through the help of regularization, the designed deep learning framework can solve the overfitting problem. The main contributions in this framework are to simulate the perception of retargeted images in HVS. Especially, it trains two separated SAE models based on geometrical shape and content matching. Then, the weighting schemes can be used to combine the obtained scores from two models. Experimental results in three well-known databases show that our method can achieve better performance than traditional methods in evaluating different image retargeting results.

8.
IEEE Trans Neural Netw Learn Syst ; 31(6): 2030-2041, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-31449032

RESUMO

Accurate measurement of position and attitude information is particularly important. Traditional measurement methods generally require high-precision measurement equipment for analysis, leading to high costs and limited applicability. Vision-based measurement schemes need to solve complex visual relationships. With the extensive development of neural networks in related fields, it has become possible to apply them to the object position and attitude. In this paper, we propose an object pose measurement scheme based on convolutional neural network and we have successfully implemented end-to-end position and attitude detection. Furthermore, to effectively expand the measurement range and reduce the number of training samples, we demonstrated the independence of objects in each dimension and proposed subadded training programs. At the same time, we generated generating image encoder to guarantee the detection performance of the training model in practical applications.

9.
Oncol Lett ; 18(6): 6323-6330, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31807157

RESUMO

Podoplanin (PDPN) is an important positive regulator of platelet aggregation and functions as a lymphatic endothelial marker. PDPN has been observed to be expressed in human tumor tissues and various cancer cell lines. In the present study, PDPN expression in patients with primary osteosarcoma was assessed at the mRNA and protein levels, and the associations between PDPN expression and pulmonary metastasis (PM) and prognosis were examined. Reverse transcription-quantitative PCR (RT-qPCR) analysis was used to detect the expression levels of PDPN in primary osteosarcoma tissues and paired normal bone tissues (n=20 pairs). In addition, immunohistochemical analysis of PDPN expression was performed in 168 paraffin-embedded osteosarcoma tissue specimens and 23 matched normal tissues. The RT-qPCR results revealed higher mRNA expression levels of PDPN in patients with PM compared with patients without PM. Further survival analyses identified Enneking stage and PM as two independent prognostic indicators. Finally, univariate analysis revealed that high PDPN protein expression was significantly associated with Enneking stage and PM in patients with osteosarcoma.

10.
Neurol Res ; 41(11): 972-979, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31296147

RESUMO

Objective: Bone cancer pain is characterized by moderate to severe ongoing pain that commonly requires the use of opiates. Transient receptor potential vanilloid subfamily member 1 (TRPV1), a new target of the analgesics, activated by heat, protons and capsaicin and the hot component of pepper. However, little is known of the anti-nociceptive effects of TRPV1 in cancer-induced bone pain. RNA interference (RNAi) has proven to be a powerful technique to study the function of genes by producing knock-down phenotypes. The aim of this study is to investigate the potential role of TRPV1 in rat model of bone cancer pain. Methods: Bone cancer pain animal model was created by tumor cell implantation (TCI). An AAV-mediated siRNA against TRPV1 was intrathecally delivered into the rats. Animal behaviors were measured using a set of mechanical or electronic von Frey apparatus and hot plate. mRNA and protein expression were examined by using qPCR and western blot methods. Results: Mechanical threshold and paw withdrawal latency in response to thermal stimulation were significantly elevated in rats with intrathecal administration of AAV-mediated siRNA against TRPV1. Moreover, class I histone deacetylases (HDACs), which plays a critical role in the neuro-inflammation response, and TNFα in the spinal cord were also significantly suppressed upon knockdown of TRPV1 by AAV-mediated siRNA against TRPV1 in rat spinal cord. Conclusions: Knockdown of TRPV1 effectively ameliorated mechanical allodynia and thermal hyperalgesia induced by TCI. Our data demonstrated that modulate the expression of TRPV1 in the spinal cord could be a potential therapeutic approach for bone cancer pain.


Assuntos
Dor do Câncer/tratamento farmacológico , Nociceptividade/efeitos dos fármacos , RNA Interferente Pequeno/farmacologia , Canais de Cátion TRPV/genética , Analgésicos Opioides/uso terapêutico , Animais , Dor do Câncer/genética , Modelos Animais de Doenças , Gânglios Espinais/metabolismo , Masculino , Interferência de RNA/fisiologia , RNA Interferente Pequeno/genética , Ratos Sprague-Dawley , Medula Espinal/patologia
11.
Sci Rep ; 9(1): 366, 2019 01 23.
Artigo em Inglês | MEDLINE | ID: mdl-30675003

RESUMO

To address the unmet needs for human polyclonal antibodies both as therapeutics and diagnostic reagents, building upon our previously established transchromosomic (Tc) cattle platform, we report herein the development of a Tc goat system expressing human polyclonal antibodies in their sera. In the Tc goat system, a human artificial chromosome (HAC) comprising the entire human immunoglobulin (Ig) gene repertoire in the germline configuration was introduced into the genetic makeup of the domestic goat. We achieved this by transferring the HAC into goat fetal fibroblast cells followed by somatic cell nuclear transfer for Tc goat production. Gene and protein expression analyses in the peripheral blood mononuclear cells (PBMC) and the sera, respectively, of Tc caprine demonstrated the successful expression of human Ig genes and antibodies. Furthermore, immunization of Tc caprine with inactivated influenza A (H7N9) viruses followed by H7N9 Hemagglutinin 1 (HA1) boosting elicited human antibodies with high neutralizing activities against H7N9 viruses in vitro. As a small ungulate, Tc caprine offers the advantages of low cost and quick establishment of herds, therefore complementing the Tc cattle platform in responses to a range of medical needs and diagnostic applications where small volumes of human antibody products are needed.


Assuntos
Anticorpos Antivirais/imunologia , Formação de Anticorpos/imunologia , Especificidade de Anticorpos/imunologia , Subtipo H7N9 do Vírus da Influenza A/imunologia , Animais , Animais Geneticamente Modificados , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/sangue , Cromossomos Artificiais Humanos , Ensaio de Imunoadsorção Enzimática , Engenharia Genética , Cabras , Humanos , Imunização , Imunoglobulina G/genética , Imunoglobulina G/imunologia , Testes de Neutralização
12.
JCI Insight ; 3(19)2018 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-30282831

RESUMO

Cystic fibrosis (CF) is a genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. The major cause of limited life span in CF patients is progressive lung disease. CF models have been generated in 4 species (mice, rats, ferrets, and pigs) to enhance our understanding of the CF pathogenesis. Sheep may be a particularly relevant animal to model CF in humans due to the similarities in lung anatomy and development in the two species. Here, we describe the generation of a sheep model for CF using CRISPR/Cas9 genome editing and somatic cell nuclear transfer (SCNT) techniques. We generated cells with CFTR gene disruption and used them for production of CFTR-/- and CFTR+/- lambs. The newborn CFTR-/- sheep developed severe disease consistent with CF pathology in humans. Of particular relevance were pancreatic fibrosis, intestinal obstruction, and absence of the vas deferens. Also, substantial liver and gallbladder disease may reflect CF liver disease that is evident in humans. The phenotype of CFTR-/- sheep suggests this large animal model will be a useful resource to advance the development of new CF therapeutics. Moreover, the generation of specific human CF disease-associated mutations in sheep may advance personalized medicine for this common genetic disorder.


Assuntos
Sistemas CRISPR-Cas/genética , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Fibrose Cística/genética , Modelos Animais de Doenças , Ovinos , Animais , Animais Geneticamente Modificados , Fibrose Cística/patologia , Feminino , Fibrose , Vesícula Biliar/patologia , Técnicas de Inativação de Genes , Humanos , Fígado/patologia , Pulmão/patologia , Masculino , Técnicas de Transferência Nuclear , Pâncreas/patologia , Fenótipo , Especificidade da Espécie
13.
Artigo em Inglês | MEDLINE | ID: mdl-30371364

RESUMO

Most of the current blind stereoscopic image quality assessment (SIQA) algorithms cannot show reliable accuracy. One reason is that they do not have the deep architectures and the other reason is that they are designed on the relatively weak biological basis, compared with findings on human visual system (HVS). In this paper, we propose a Deep Edge and COlor Signal INtegrity Evaluator (DECOSINE) based on the whole visual perception route from eyes to the frontal lobe, and especially focus on edge and color signal processing in retinal ganglion cells (RGC) and lateral geniculate nucleus (LGN). Furthermore, to model the complex and deep structure of the visual cortex, Segmented Stacked Auto-encoder (S-SAE) is used, which has not utilized for SIQA before. The utilization of the S-SAE complements weakness of deep learning-based SIQA metrics that require a very long training time. Experiments are conducted on popular SIQA databases, and the superiority of DECOSINE in terms of prediction accuracy and monotonicity is proved. The experimental results show that our model about the whole visual perception route and utilization of S-SAE are effective for SIQA.

14.
Med Sci Monit ; 24: 4926-4933, 2018 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-30008466

RESUMO

BACKGROUND The aim of this study was investigate the effects of the sesquiterpene lactone, ludartin, on cell proliferation, cell migration, apoptosis, and the cell cycle in osteosarcoma cell lines, compared with a normal osteoblast cell line. MATERIAL AND METHODS Osteosarcoma cell lines, MG-63 Saos-2 U-2OS, T1-73 143B, and HOS, and normal hFOB 1.19 osteoblasts, were cultured and treated with increasing doses of ludartin, The MTT colorimetric assay was used to measure cell metabolic activity and viability. Apoptosis was studied by fluorescence-activated cell sorting (FACS) using 4',6-diamidino-2-phenylindole (DAPI) nuclear staining and Annexin-V/propidium iodide (PI) staining. Cell cycle was studied using flow cytometry. Cell migration and invasion were studied using wound healing and Boyden chamber assays. Protein expression was measured by Western blotting. RESULTS Ludartin inhibited cell viability, cell migration, cell proliferation, and increased cell apoptosis, in all osteosarcoma cell lines, with an IC50 dose ranging from 15-30 µM. The greatest effects were on the Saso-2 osteosarcoma cells, with an IC50 of 15 µM. However, ludartin showed minor cytotoxic effects of the normal hFOB 1.19 osteoblasts (IC50 >100 µM). Ludartin exerted its anti-proliferative effects on Saos-2 cells via induction of apoptosis and cell cycle arrest at the G2/M checkpoint, associated with reduced expression of Cdc25c (Ser216), Cdc25c, pCdc2 (Tyr15), and Cdc2 and increased expression of p21WAF1. Ludartin inhibited cell migration and invasion of the Saos-2 cells. CONCLUSIONS The dose-dependent effects of ludartin on cell proliferation, migration, apoptosis, cell cycle arrest at the G2/M checkpoint involved p21WAFI in Saos-2 osteosarcoma cells.


Assuntos
Neoplasias Ósseas/tratamento farmacológico , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Osteossarcoma/tratamento farmacológico , Sesquiterpenos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Ciclo Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Osteoblastos/metabolismo , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Ativação Transcricional , Regulação para Cima/efeitos dos fármacos
15.
Oncol Lett ; 16(2): 2305-2311, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30008933

RESUMO

Previous studies have demonstrated that connective tissue growth factor (CTGF) is expressed at increased levels in prostate cancer bone metastasis mouse models and patients with prostate cancer which metastasizes to the bone; however, the underlying molecular mechanism(s) remain unknown. The present study investigated the function of CTGF in osteoblast differentiation and its effect on prostate cancer bone metastasis by analyzing CTGF gene expression and transcription at different levels of invasion, metastasis of prostate cancer cells, and the influence of CTGF on proliferation and xenotransplantation. A mouse model demonstrating bone metastasis was used to investigate the function(s) of CTGF in bone metastasis and osteoblast differentiation. Results demonstrated that CTGF expression was increased in association with high bone metastasis in prostate cancer cells, and its expression was significantly decreased in whole cell lysates. CTGF expression in prostate cancer cells with high levels of bone metastasis was increased 1.9-fold compared with prostate cancer cells with low levels of bone metastasis. The expression of CTGF in mesenchymal cells was markedly increased compared with epithelial cells. Results indicated that the increased expression of CTGF does not affect the proliferation of tumor cells and possesses no influence on tumor volume. Control and CTGF plasmids were transfected into RM1 cells and led to 4 and 17% bone lesions, respectively. Increased expression of CTGF significantly enlarged the tumor area in the bone metastatic position compared with the control. Positive areas of alkaline phosphatase were significantly decreased as the concentration of CTGF increased. The results of the present study demonstrated that CTGF promotes prostate carcinoma to metastasize in the bone by dysregulating osteoblast differentiation.

16.
Medicine (Baltimore) ; 97(27): e11418, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29979441

RESUMO

This retrospective study investigated the effect and safety of extracorporeal shockwave therapy (ESWT) for treatment of knee osteoarthritis (KOA).In this retrospective study, 105 patients with KOA were included. Of those, 60 patients underwent ESWT, whereas 45 patients received laser therapy. Effect was measured by the Numeric Rating Scale (NRS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). In addition, adverse events (AEs) were also recorded in this study. All outcome measurements were evaluated at the end of weeks 1, 6, and 12.Compared with the laser therapy, ESWT showed greater effect in KOA symptoms relief with regard to NRS, WOMAC total, and its subscores at week 6 (P < .05) and week 12 (P < .01) after treatment. No AE, however, occurred in both groups.The results of this retrospective study found that ESWT may be efficacious and safe for the treatment of patients with KOA. It, however, had an intrinsic limitation as a retrospective study. Prospective study with larger sample size is still needed to warrant the result of this study in the future.


Assuntos
Tratamento por Ondas de Choque Extracorpóreas/métodos , Terapia a Laser/métodos , Osteoartrite do Joelho/terapia , Idoso , Artralgia/terapia , Feminino , Humanos , Articulação do Joelho , Masculino , Pessoa de Meia-Idade , Medição da Dor , Estudos Retrospectivos , Índice de Gravidade de Doença , Resultado do Tratamento
17.
Cell Reprogram ; 20(3): 187-195, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29688743

RESUMO

Unlike oocytes of many other mammalian species, parthenogenetically activated hamster oocytes have not been reported to develop beyond the two-cell stage. This study investigated the in vitro development into blastocysts of parthenogenetic embryos of Golden Syrian hamsters. We observed that hamster oocytes could easily be artificially activated (AA) by treatment with ionomycin plus 6-dimethylaminopurine + cycloheximide + cytochalasin B as assessed by embryo cleavage in HECM-9 (63.15%) or HECM-10 (63.82%). None of the cleaved embryos developed beyond the two-cell stage when cultured in either of the two media. However, some of the embryos overcame the two-cell block and developed to the blastocyst stage (26.45%) when they were first cultured in HECM-10 for 24 hours and then in HECM-9 (serial culture media HECM-10-9) for 72 hours. Blastocyst development was further significantly (66.2%) improved when embryos were cultured in HECM-10 supplemented with ethylenediaminetetraacetic acid for 24 hours, then in HECM-9 supplemented with glucose for 72 hours (serial culture media HECM-11a-b). Hamster oocytes activated with ionomycin, ethanol, or a combination of the two treatments would develop to the blastocyst stage in serial culture media HECM-11a-b, whereas none of the spontaneously activated oocytes cleaved (0% vs. 86.93%, p < 0.05). DNA and microtubule configurations of spontaneously activated and AA oocytes were assessed by immunocytochemical staining and fluorescence microscopy. The results indicate that serial culture and the method of activation are critical for overcoming the in vitro developmental block of hamster parthenogenetic embryos. This study is the first to report blastocyst development from parthenogenetically activated hamster oocytes.


Assuntos
Blastocisto/fisiologia , Técnicas de Cultura Embrionária/veterinária , Oócitos/fisiologia , Partenogênese , Adenina/administração & dosagem , Adenina/análogos & derivados , Animais , Blastocisto/efeitos dos fármacos , Meios de Cultura , Cicloeximida/administração & dosagem , Citocalasina B/administração & dosagem , Técnicas de Cultura Embrionária/métodos , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Ionomicina/administração & dosagem , Mesocricetus , Oócitos/efeitos dos fármacos
18.
Int J Mol Med ; 41(6): 3680-3690, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29568936

RESUMO

The aim of this study was to examine the effects of the insulin gene enhancer-binding protein, islet-1 (ISL1), on the proliferation, invasion and apoptosis of the human melanoma cell line, A375. An ISL1 overexpression lentiviral vector was constructed and transfected into the A375 cells. The proliferation of the A375 cells transfected with the ISL1 vector (termed A375/ISL1 cells) was examined by MTT assay, flow cytometry and TUNEL assay, and cell invasion was examined by Transwell assay. The expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 were measured by qPCR and western blot analysis; the expression levels of Akt and p-Akt were measured in the cells treated with vascular endothelial growth factor (VEGF) and the PI3K/Akt inhibitor, LY294002, by western blot analysis. The optical density value of the A375/ISL1 cells was increased after 12 h of culture (P<0.001), as shown by MTT assay. The ratio of apoptotic A375/ISL1 cells was significantly decreased (P<0.001), as shown by flow cytometry and TUNEL assay. In addition, the average penetration rate of the A375/ISL1 cells significantly increased (P<0.001), as shown by Transwell assay. The expression levels of MMP-2 and MMP-9 were significantly increased in the A375/ISL1 cells, as shown by qPCR and western blot analysis (P<0.001). Moreover, treatment of the A375/ISL1 cells with VEGF for 48 h increased the expression of Akt and p-Akt compared with the control cells transfected with A375/green fluorescent protein (GFP) (P<0.05; P<0.001, respectively). In addition, in the A375/ISL1 cells treated with the LY294002 inhibitor for 24 and 48 h, the level of Akt was also found to increase compared to the control A375/GFP cells (P<0.05). On the whole, the findings of this study indicate that the overexpression of ISL1 promotes the proliferation and invasion, and inhibits the apoptosis of A375 melanoma cells. ISL1 thus plays an important role in A375 cell survival, and these effects are possibly mediate via the PI3K/Akt signaling pathway.


Assuntos
Melanoma/metabolismo , Apoptose/genética , Apoptose/fisiologia , Ciclo Celular/genética , Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Proliferação de Células/fisiologia , Citometria de Fluxo , Regulação Neoplásica da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/fisiologia , Humanos , Marcação In Situ das Extremidades Cortadas , Melanoma/patologia , Transdução de Sinais/genética , Transdução de Sinais/fisiologia
19.
Am J Reprod Immunol ; 79(1)2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29131441

RESUMO

PROBLEM: The regulatory mechanisms governing differential expression of classical major histocompatibility complex (MHC) class I (MHC-Ia) and non-classical MHC class I (MHC-Ib) genes are poorly understood. METHOD OF STUDY: Quantitative reverse transcription- polymerase chain reaction (PCR) was used to compare the abundance of MHC-I transcripts and related transcription factors in peripheral blood mononuclear cells (PBMC) and placental trophoblast cells (PTC). Methylation of MHC-I CpG islands was detected by bisulfite treatment and next-generation sequencing. Demethylation of PBMC and PTC with 5'-aza-deoxycytidine was used to assess the role of methylation in gene regulation. RESULTS: MHC-I expression was higher in PBMC than PTC and was correlated with expression of IRF1, class II MHC transactivator (CIITA), and STAT1. The MHC-Ia genes and BoLA-NC1 were devoid of CpG methylation in PBMC and PTC. In contrast, CpG sites in the gene body of BoLA-NC2, -NC3, and -NC4 were highly methylated in PBMC but largely unmethylated in normal PTC and moderately methylated in somatic cell nuclear transfer PTC. In PBMC, demethylation resulted in upregulation of MHC-Ib by 2.8- to 6-fold, whereas MHC-Ia transcripts were elevated less than 2-fold. CONCLUSION: DNA methylation regulates bovine MHC-Ib expression and is likely responsible for the different relative levels of MHC-Ib to MHC-Ia transcripts in PBMC and PTC.


Assuntos
Bovinos , Antígenos de Histocompatibilidade Classe I/metabolismo , Leucócitos Mononucleares/fisiologia , Placenta/fisiologia , Trofoblastos/fisiologia , Animais , Células Cultivadas , Ilhas de CpG/genética , Metilação de DNA , Epigênese Genética , Feminino , Regulação da Expressão Gênica , Antígenos de Histocompatibilidade Classe I/genética , Fator Regulador 1 de Interferon/metabolismo , Gravidez , Fator de Transcrição STAT1/metabolismo
20.
IEEE Trans Cybern ; 48(9): 2583-2597, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28976326

RESUMO

This paper considers the problem of maximizing the number of task allocations in a distributed multirobot system under strict time constraints, where other optimization objectives need also be considered. It builds upon existing distributed task allocation algorithms, extending them with a novel method for maximizing the number of task assignments. The fundamental idea is that a task assignment to a robot has a high cost if its reassignment to another robot creates a feasible time slot for unallocated tasks. Multiple reassignments among networked robots may be required to create a feasible time slot and an upper limit to this number of reassignments can be adjusted according to performance requirements. A simulated rescue scenario with task deadlines and fuel limits is used to demonstrate the performance of the proposed method compared with existing methods, the consensus-based bundle algorithm and the performance impact (PI) algorithm. Starting from existing (PI-generated) solutions, results show up to a 20% increase in task allocations using the proposed method.

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