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1.
J Cell Physiol ; 2020 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-31951022

RESUMO

Traumatic osteonecrosis of femoral head (TONFH) is a common orthopedic disease caused by physical injury in hip. However, the unclear pathogenesis mechanism of TONFH and lacking of simple noninvasive early diagnosis method cause the necessity of hip replacement for most patients with TONFH. In this study, we aimed to identify circulating microRNAs (miRNAs) by integrated bioinformatics analyses as potential biomarker of TONFH. mRNA expression profiles were downloaded from the Gene Expression Omnibus database. Then we combined two miRNA screen methods: Weighted gene co-expression network analysis and fold change based differentially expressed miRNAs analysis. As a result, we identified 14 key miRNAs as potential biomarkers for TONFH. Besides, 302 target genes of these miRNAs were obtained and the miRNA-mRNA interaction network was constructed. Furthermore, the results of Kyoto Encyclopedia of Gene and Genome pathway analysis, Gene Ontology function analysis, protein-protein interaction (PPI) network analysis and PPI network module analysis showed close correlation between these 14 key miRNAs and TONFH. Then we established receiver operating characteristic curves and identified 6-miRNA signature with highly diagnosis value including miR-93-5p (area under the curve [AUC] = 0.93), miR-1324 (AUC = 0.92), miR-4666a-3p (AUC = 0.92), miR-5011-3p (AUC = 0.92), and miR-320a (AUC = 0.89), miR-185-5p (AUC = 0.89). Finally, the results of quantitative real-time polymerase chain reaction confirmed the significantly higher expression of miR-93-5p and miR-320a in the serum of patients with ONFH. These circulating miRNAs could serve as candidate early diagnosis markers and potential treatment targets of TONFH.

2.
Mol Cell Probes ; 49: 101474, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31655106

RESUMO

The SYBR Green І-based duplex real-time PCR assay was developed for simultaneous detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 3 (PCV-3) genomes. PRRSV and PCV-3 were distinguished in the same sample by their distinctive melting temperature (Tm) which was 84 °C for PRRSV and 81.5 °C for PCV-3, and other non-targeted swine viruses showed no specific melting peaks. The detection limits of this assay were 46.1copies/µL for PRRSV and 49.3copies/µL for PCV-3, respectively. Thirty-three lung samples of porcine with respiratory and reproductive failure symptoms were collected and confirmed by the SYBR Green І-based real-time PCR assay and conventional PCR assay. The real-time PCR detection results showed that the PRRSV positive rate was 45.45%, the PCV-3 positive rate was 63.63%, the PRRSV and PCV-3 co-infection positive rate was 36.36%, which were more sensitive than conventional PCR detection. This duplex real-time PCR assay could be a rapid, sensitive and reliable method for the detection of PRRSV and PCV-3 co-infection.

3.
Sci Total Environ ; 703: 135562, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-31767303

RESUMO

Plant-soil systems have complex regulatory mechanisms for xenobiotics uptake by plant, and these chemicals in soil pore water (SPW) are regarded as the bioavailable fraction. To date, little is known about the role of SPW in regard to the bioavailability of antibiotics for plant. In this study, in situ soil moisture sampler (SMS) was adopted to collect SPW from four paddy soils without disrupting the rhizosphere zone to evaluate antibiotic uptake in rice. The results show that SMS is applicable for antibiotics that have small molecular sizes and Log Kow values, e.g., sulfadiazine (SDZ), sulfamethoxazole (SMZ), trimethoprim (TRM), and florfenicol (FLR). However, SMS performance was not feasible for large size and medium hydrophobic clarithromycin (CLR). Antibiotics in SPW demonstrated differences among chemicals and soils. Relatively higher levels of SDZ, SMZ, and FLR were observed in SPW than TRM, and neutral Panjin soil had the highest levels of antibiotics in SPW among four soils. The levels of antibiotics in SPW were negatively correlated with their soil partition parameter, Kd. Rapid decreases of SMZ, FLR, and SDZ in the SPW were consistent with their low residues in the final soils. All antibiotics were detectable in rice roots, followed by low detection levels in a few shoot samples, while no antibiotics were detectable in the grains of four soils. Relatively higher levels of SDZ, SMZ and FLR were observed in the roots of Panjin soil, consistent with their levels in SPW and Kd values. Furthermore, CLR and TRM were observed to have higher levels in roots, which was regarded as a consequence of their relatively longer persistence. Our study indicates that SMS is an applicable technique for in situ sampling of SPW, and level of antibiotics in SPW can work as a useful indicator to explore their bioavailability to plants.


Assuntos
Antibacterianos/análise , Monitoramento Ambiental , Poluentes do Solo/análise , Rizosfera , Solo/química , Sulfametoxazol , Trimetoprima , Água
4.
Artigo em Inglês | MEDLINE | ID: mdl-31574957

RESUMO

Adolescents engage in health risk behaviors (HRBs) that influence their current and future health status. Health literacy (HL) is defined as how well a person can get and understand the health information and services, and use them to make good health decisions. HL can be used to participate in everyday activities actively and apply new information to the changing circumstances. HRBs commonly co-occur in adolescence, and few researchers have examined how HL predicts multiple HRBs in adolescence. In this study we examined the subgroups of HRBs, and investigated heterogeneity in the effects of HL on the subgroups. In total, 22,628 middle school students (10,990 males and 11,638 females) in six cities were enrolled by multistage stratified cluster sampling from November 2015 to January 2016. The measurement of HL was based on the Chinese Adolescent Interactive Health Literacy Questionnaire (CAIHLQ). Analyses were conducted with regression mixture modeling approach (RMM) by Mplus. By this study we found four latent classes among Chinese adolescents: Low-risk class, moderate-risk class 1 (smoking/alcohol use (AU)/screen time (ST)), moderate-risk class 2 (non-suicidal self-injury (NSSI)/suicidal behaviors (SB)/unintentional injury (UI)), and high-risk class (smoking/AU/ST/NSSI/SB/UI) which were 64.0%, 4.5%, 28.8% and 2.7% of involved students, respectively. Negative correlations were found between HL and HRBs: higher HL accompanied decreased HBRs. Compared to the low-risk class, moderate-risk class 1 (smoking/AU/ST), moderate-risk class 2 (NSSI/SB/UI), and high-risk class (smoking/AU/ST/NSSI/SB/UI) showed OR (95%CI) values of 0.990 (0.982-0.998), 0.981 (0.979-0.983) and 0.965 (0.959-0.970), respectively. Moreover, there was heterogeneity in the profiles of HRBs and HL in different classes. It is important for practitioners to examine HRBs in multiple domains concurrently rather than individually in isolation. Interventions and research should not only target adolescents engaging in high levels of risky behavior but also adolescents who are engaging in lower levels of risky behavior.


Assuntos
Grupo com Ancestrais do Continente Asiático , Alfabetização em Saúde , Comportamentos de Risco à Saúde , Adolescente , China , Cidades , Coleta de Dados , Feminino , Humanos , Masculino , Fumar , Inquéritos e Questionários , Adulto Jovem
5.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 27(5): 1469-1475, 2019 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-31607300

RESUMO

OBJECTIVE: To investigate the effect of LNK gene silencing and overexpression on the expression of STAT3 gene in human monocytic leukemia cells (THP-1). METHODS: THP-1 cells were cultured, and the lentivirus was used as a vector to silence and overexpres the LNK gene stably. After transfection for 72 hours, the GFP expression levels were observed by inverted fluorescence microscopy. The lentiviral transfection efficiencies were detected by flow cytometry. The effects of LNK silencing and overexpression were confirmed, and the expression of STAT3 mRNA was detected by RT-PCR. The protein levels of LNK and STAT3 were detected by Western blotting. RESULTS: The GFP expression level of THP-1 cells reached more than 85% after transfection with lentivirus for 72 hours, and the transfection efficiency of cells was above 99%. mRNA expressions levels of LNK and STAT3 in LNK silencing group were signifycantly lower than those in control group, while LNK and STAT3 mRNA levels in the LNK overexpression group was significantly higher than those in control group (P<0.05). The protein expression levels of LNK and STAT3 in LNK silencing group were significantly lower than those in control group, while that in LNK overexpression group was significantly higher than that in control group (P<0.05). CONCLUSION: The THP-1 cell line with LNK gene silencing and overexpression has been successfully established. The LNK gene silencing resulted in decrease of STAT3 expression; LNK gene overexpression and leads to inereases of STAT3 expression indicating that LNK participates in the regulation of STAT3.


Assuntos
Fator de Transcrição STAT3/metabolismo , Linhagem Celular Tumoral , Inativação Gênica , Vetores Genéticos , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Lentivirus , Proteínas , RNA Interferente Pequeno , Células THP-1 , Transfecção
6.
Nano Lett ; 19(11): 8049-8058, 2019 11 13.
Artigo em Inglês | MEDLINE | ID: mdl-31558023

RESUMO

Pyroptosis is a lytic and inflammatory form of programmed cell death and could be induced by chemotherapy drugs via caspase-3 mediation. However, the key protein gasdermin E (GSDME, translated by the DFNA5 gene) during the caspase-3-mediated pyroptosis process is absent in most tumor cells because of the hypermethylation of DFNA5 (deafness autosomal dominant 5) gene. Here, we develop a strategy of combining decitabine (DAC) with chemotherapy nanodrugs to trigger pyroptosis of tumor cells by epigenetics, further enhancing the immunological effect of chemotherapy. DAC is pre-performed with specific tumor-bearing mice for demethylation of the DFNA5 gene in tumor cells. Subsequently, a commonly used tumor-targeting nanoliposome loaded with cisplatin (LipoDDP) is used to administrate drugs for activating the caspase-3 pathway in tumor cells and trigger pyroptosis. Experiments demonstrate that the reversal of GSDME silencing in tumor cells is achieved and facilitates the occurrence of pyroptosis. According to the anti-tumor activities, anti-metastasis results, and inhibition of recurrence, this pyroptosis-based chemotherapy strategy enhances immunological effects of chemotherapy and also provides an important insight into tumor immunotherapy.

7.
Brain Res ; 1725: 146472, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31545956

RESUMO

Patients suffering from depression most commonly present with symptoms associated with the autonomic nervous system. Despite the satisfactory results achieved following treatment with vagus nerve stimulation and drug treatment, recurrence is a common occurrence in many patients. As described in numerous studies, prolactin receptor (PRLR) has been identified as an anxiolytic and anti-depressant factor in depression. However, the effect of PRLR on chronic mild stress (CMS)-induced depression remains to be thoroughly demonstrated. Therefore, the present study was conducted on the effect of PRLR gene on brain derived neurotrophic factor (BDNF) expression and hippocampal neuron apoptosis through the establishment of CMS-induced depression mouse models, with aims of providing a new and effective therapeutic option for depression. Microarray-based analysis was initially used to retrieve depression-related expression dataset and PRLR-related signaling pathway. Lentiviral vectors overexpressing PRLR or expressing PRLR-specific shRNA were used to up- or down-regulated the expression of PRLR in mice. Subsequently, the effects of PRLR on hippocampal neurons and pyramidal cells in CA1 and CA3 regions, and ultrastructure in hippocampal region were evaluated. Serum BDNF level and the positive rate of cleaved-Caspase-3 in hippocampal CA3 region were determined. Next, the regulatory mechanism by which PRLR gene silencing influences hippocampal neuron apoptosis via the JAK2-STAT5 signaling pathway was detected. PRLR gene was assumed to participate in the development of depression by regulating the JAK-STAT signaling pathway. Our results found that the mice with CMS-induced depression exhibited locomotion activity and anhedonia. In addition, a decrease in the number of pyramidal cells was observed in the hippocampus while that of apoptotic cells was increased. In addition, serum BDNF level was increased, and the expression of Caspase-3 and Bax in hippocampal neurons and the JAK2-STAT5 signaling pathway was decreased in response to PRLR silencing, along with increased expression of BDNF and Bcl-2. From the aforementioned findings, we concluded that PRLR gene silencing results in the inhibition of hippocampal neuron apoptosis and alleviation of CMS-induced depression by inactivating the JAK2-STAT5 signaling pathway and elevating BDNF expression, providing a new insight for the treatment of depression.

8.
J Transl Med ; 17(1): 256, 2019 08 07.
Artigo em Inglês | MEDLINE | ID: mdl-31391086

RESUMO

BACKGROUND: The adipose afferent reflex (AAR), a sympatho-excitatory reflex, can promote the elevation of sympathetic nerve activity (SNA) and blood pressure (BP). Inflammation in the paraventricular nucleus (PVN) involves sympathetic abnormality in some cardiovascular diseases such as hypertension. This study was designed to explore the effects of tumor necrosis factor alpha (TNFα) in the PVN on the AAR and SNA in rats with obesity-related hypertension (OH) induced by a high-fat diet for 12 weeks. METHODS: Renal sympathetic nerve activity (RSNA) and mean arterial pressure (MAP) were continuously recorded in anesthetized rats, and their responses to capsaicin (CAP) stimulation of the right inguinal white adipose tissue were used to evaluate the AAR. RESULTS: Compared to the control rats, the systolic blood pressure (SBP), plasma norepinephrine (NE, indicating SNA) and TNFα levels, TNFα mRNA and protein levels, reactive oxygen species (ROS) content and NADPH oxidase activity in the PVN were significantly elevated in rats with OH. TNFα in the PVN markedly enhanced sympathoexcitation and AAR. Moreover, the enhancement of AAR caused by TNFα can be significantly strengthened by the pretreatment of diethyldithiocarbamate (DETC), a superoxide dismutase inhibitor, but attenuated by TNF-α receptor antagonist R-7050, superoxide scavenger PEG-SOD and NADPH oxidase inhibitor apocynin (Apo) in rats with OH. Acute microinjection of TNF-α into the PVN significantly increased the activity of NADPH oxidase and ROS levels in rats with OH, which were effectively blocked by R-7050. Furthermore, our results also showed that the increased levels of ROS, TNFα and NADPH oxidase subunits mRNA and protein in the PVN of rats with OH were significantly reversed by pentoxifylline (PTX, 30 mg/kg daily ip; in 10% ethanol) application, a cytokine blocker, for a period of 5 weeks. PTX administration also significantly decreased SBP, AAR and plasma NE levels in rats with OH. CONCLUSIONS: TNFα in the PVN modulates AAR and contributes to sympathoexcitation in OH possibly through NADPH oxidase-dependent ROS generation. TNFα blockade attenuates AAR and sympathoexcitation that unveils TNFα in the PVN may be a possible therapeutic target for the intervention of OH.

9.
Angew Chem Int Ed Engl ; 58(40): 14224-14228, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31389144

RESUMO

The targeted delivery of chemotherapeutic drugs is a major challenge in the clinical treatment of cancer. Herein, we constructed a multifunctional DNA nanoplatform as a versatile carrier of the highly potent platinum-based DNA intercalator, 56MESS. In our rational design, 56MESS was efficiently loaded into the double-bundle DNA tetrahedron through intercalation with the DNA duplex. With the integration of a nanobody that both targets and blocks epidermal growth factor receptor (EGFR), the DNA nanocarriers exhibit excellent selectivity for cells with elevated EGFR expression (a common biomarker related to tumor formation) and combined tumor therapy without obvious systemic toxicity. This DNA-based platinum-drug delivery system provides a promising strategy for the treatment of tumors.

10.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 27(4): 1013-1019, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31418350

RESUMO

OBJECTIVE: To investigate the effect of silencing LNK gene on the expression of EPO and EPOR in acute myeloid leukemia cells (THP-1). METHODS: THP-1 cells were cultured. The lentivirus was used as a vector to silence the LNK gene stably. After 72 hours of infection, GFP expression level was detected by the fluorescent inverted microscopy. The lentiviral Infection efficiencies were monitored by flow cytometry. The LNK silencing effect was confirmed. The mRNA expressions of EPO and EPOR were detected by RT-PCR. The protein levels of LNK, EPO and EPOR were detected by Western blot. RESULTS: At the time-point of 72 hours after lentivirus infection, the expression level of GFP was above 85% detected by fluorescent inverted microscopy. The infection efficiency was above 99% by flow cytometry. mRNA expressions of LNK, EPO and EPOR in LNK silencing group were signifycantly lower than those in control group (P<0.05). The protein levels of LNK, EPO and EPOR in LNK silencing group were significantly lower than those in the control group (P<0.05). CONCLUSION: THP-1 cell line of LNK gene silencing has been successfully established,the LNK gene has been silenced, the expression of EPO and EPOR decrease, indicating that LNK may participate in the regulation of EPO and EPOR.


Assuntos
Proteínas/genética , Western Blotting , Eritropoetina , Inativação Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Receptores da Eritropoetina , Células THP-1
11.
Zhongguo Shi Yan Xue Ye Xue Za Zhi ; 27(4): 1026-1032, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31418352

RESUMO

OBJECTIVE: To investigate the expression of erythropoietin (EPO) and erythropoietin receptor (EPOR) in patients with acute leukemia (AL) and its clinical significance. METHODS: The levels of EPO and EPOR in plasma were determined by ELISA kit. mRNA expression levels of EPO and EPOR were determined by RT-RCR. The protein expression levels of EPO and EPOR were detected by Western blot. RESULTS: The EPO protein levels in marrow plasma of ALL and AML group were significantly higher than those in the control group (P<0.05), EPOR protein levels in ALL and AML group were significantly lower than those in the control group (P<0.05). The mRNA levels of EPO and EPOR in ALL and AML groups were significantly higher than those in the control group (P<0.05). The mRNA levels of EPO and EPOR in the high risk ALL and AML groups were significantly higher than those in the medium, low risk group and the control group (P<0.05). The protein expression levels of EPO and EPOR in ALL and AML groups were significantly higher than that in control group (P<0.05). The mRNA levels of EPO and EPOR in ALL and AML groups did not correlate with hemoglobin level and erythrocyte count (P>0.05). CONCLUSION: The expressions of EPO and EPOR is higher in ALL and AML patients. The expression levels of EPO and EPOR relate with the risk of ALL and AML. High risk patients have higher expression levels of EPO and EPOR, however, the expression levels of EPO and EPOR do not correlate with hemoglobin level and erythrocyte counting.


Assuntos
Leucemia Mieloide Aguda , Medula Óssea , Eritropoetina , Expressão Gênica , Humanos , Receptores da Eritropoetina
12.
Jpn J Infect Dis ; 72(6): 394-398, 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31257240

RESUMO

We evaluated two currently available rapid antigen detection tests (RADTs) for Respiratory syncytial virus (RSV), Sofia® RSV FIA and BinaxNOW RSV Card (BinaxNOW). Between November 2017 and February 2018, 395 nasopharyngeal swabs were collected from children diagnosed with acute respiratory infections. The swabs were evaluated using the aforementioned RADTs, the reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR), and the direct immunofluorescence assay (DFA). The sensitivity of Sofia® RSV FIA (80.82%) was significantly higher than that of BinaxNOW (53.42%) when RT-qPCR was used as the standard. This was confirmed with DFA. The sensitivities of Sofia® RSV FIA (85.4% [41/48]) and BinaxNOW (58.3% [28/48]) were higher for RSV A than for RSV B (69.6% [16/23] and 43.5% [10/23], respectively). The optimal critical cycle threshold (Ct) values on RT-qPCR that correlated with Sofia® RSV FIA and BinaxNOW were 24 and 22, respectively. The kappa value for Sofia® RSV FIA and RT-qPCR was 0.962 in patients who were two years old or younger, but 0.648 in those who were more than two years old. Thus, Sofia® RSV FIA is more sensitive than BinaxNOW; its results were affected by the RSV viral strain and load. Sofia® RSV FIA is more effective in children who are ≤ 2 years old than in those who are > 2 years old.

13.
J Cell Biochem ; 120(12): 19891-19901, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31338874

RESUMO

By differentiating into and the balance being regulated between M1 (pro-inflammatory) and M2 (anti-inflammatory) heterogeneous populations, macrophages play critical roles during the host immune response in various physiological contexts in both health and diseases. Besides regulating innate and adaptive immune capacity, macrophages are also decisively involved in tissue homeostasis. However, how resident macrophages are regulated after tissue damages is still far from elucidation. In the present study, we found that adipose-derived stem cells (ADSCs) apparently promote bone defect rehabilitation in vivo via skewing differentiation of bone marrow-derived macrophage (BMDMs) towards anti-inflammatory M2 macrophages. In vitro data demonstrated that although ADSCs have the potential to differentiate to osteoblasts and adipose cells by using standard tissue culture-differentiating conditions, these mesenchymal progenitors are mainly regulated to differentiate into osteoblasts with overexpressed runt-related transcription factor 2, osteoprotegerin, osterix, and downregulated receptor activator of nuclear factor κB ligand in the presence of BMDMs-conditioned medium. Whereas BMDMs are polarized toward M2 macrophages with higher levels of arginase 1 and mannose receptor, but lower levels of inducible nitric oxide synthase and tumor necrosis factor-α when cocultured with ADSCs. In short, all these findings collectively demonstrated that ADSCs and resident host cells can synergistically contribute to the bony repair through mutual regulation of their differentiation and cytokine secretion.

14.
Biochem Cell Biol ; 2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31075210

RESUMO

Osteosarcoma remains fatal in adolescents and young adults, with a 5-year survival rate of less than 20%. However, the detailed mechanism that regulates osteosarcoma metastasis is poorly understood. We analyzed the expression levels of miR-211-5p in clinical osteosarcoma samples and cell lines, which revealed that miR-211-5p expression was reduced in osteosarcoma. Moreover, induction of miR-211-5p in several osteosarcoma cell lines dramatically inhibited their migration and invasion capacity. Furthermore, miR-211-5p overexpression led to a significant increase of osteosarcoma cell apoptosis. Importantly, our in vivo xenograft experiments showed that miR-211-5p strongly inhibits tumorigenesis. Additionally, functional experiments demonstrated that miR-211-5p suppresses the expression of proline-rich protein 11 (PRR11) by directly binding to the 3' region of PRR11 mRNA. Moreover, we showed that PRR11 overexpression attenuated the increase of apoptosis and decreased migration and invasion when the upstream miR-211-5p was overexpressed. Our data provide new insights into the mechanisms that regulate osteosarcoma metastasis and novel potential pharmaceutical targets for personalized medicine.

15.
Mol Cell Probes ; 45: 31-36, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30980890

RESUMO

Porcine circovirus 3 (PCV3), as a newly emerged circovirus, is widely distributed in pig populations worldwide. Co-infection of PCV2 and PCV3 has been reported frequently in clinical samples. In the present study, a TB Green II-based duplex real-time polymerase chain reaction (qPCR) was developed to rapidly and differentially detect PCV2 and PCV3. The assay specifically detected PCV2 and PCV3, with no fluorescence signals being detected for other non-targeted pig pathogens. The duplex qPCR showed a high degree of linearity (R2 > 0.998), and its limits of detection were 10 and 78 copies/µL for PCV2 and PCV3, respectively. The duplex qPCR could detect and differentiate PCV2 (melting peaks at 85.5 °C) and PCV3 (melting peaks at 82.5 °C), and showed high repeatability and reproducibility, with intra- and inter-assay coefficients of variation of less than 2.0%. Fifty-six tissue samples from 18 pig farms were used to evaluate the duplex qPCR method. The results revealed infection rates of 66.07% (37/56) and 39.28% (22/56) for PCV2 and PCV3, respectively. The PCV2 + PCV3 co-infection rate was 39.28% (22/56). The developed method could be used as an efficient molecular biology tool for epidemiological investigations of PCV2 and PCV3.

16.
ACS Appl Mater Interfaces ; 11(12): 11112-11118, 2019 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-30874429

RESUMO

Delivery of proteins to carry out desired biological functions is a direct approach for disease treatment. However, protein therapy is still facing challenges due to low delivery efficiency, poor targeting during trafficking, insufficient therapeutic efficacy, and possible toxicity induced by carriers. Here, we present a novel delivery platform based on DNA origami nanostructure that enables tumor cell transportation of active proteins for cancer therapy. In our design, cytotoxic protein ribonuclease (RNase) A molecules are organized on the rectangular DNA origami nanosheets, which work as nanovehicles to deliver RNase A molecules into the cytoplasm and execute their cell-killing function inside the tumor cells. Cancer cell-targeting aptamers are also integrated onto the DNA origami-based nanoplatform to enhance its targeting effect. This DNA origami-protein coassembling strategy can be further developed to transport other functional proteins and therapeutic components simultaneously for synergistic effects and be adapted for integrated diagnostics and therapeutics.


Assuntos
DNA/química , Ribonuclease Pancreático/metabolismo , Aptâmeros de Nucleotídeos/química , Linhagem Celular Tumoral , Portadores de Fármacos/química , Humanos , Microscopia de Força Atômica , Mucina-1/química , Mucina-1/metabolismo , Nanoestruturas/química , Ribonuclease Pancreático/química
17.
Arch Virol ; 164(5): 1445-1451, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30888560

RESUMO

In order to investigate the genetic diversity of porcine circovirus type 2 (PCV2), 284 clinical tissue samples were collected from different pig farms in central China from 2015 to 2017. A total of 162 tissue samples (162/284, 57.04%) were positive for PCV2 by PCR, and subsequently, the complete genome of 36 of these PCV2 samples was cloned and sequenced. The sequencing results showed that 37 complete PCV2 sequences were obtained from 36 PCV2-positive clinical samples. These PCV2 strains were relatively conserved and extremely homologous to the representative classical PCV2 strains. Of these, 20 PCV2 strains belonged to genotype PCV2d, 14 belonged to PCV2b, and three others belonged to PCV2a. Coinfection with PCV2b and PCV2d was identified in one sample (DF-2). These results show that PCV2d may be gradually replacing PCV2b as the predominant PCV2 genotype in central China, and that other genotypes also exist in individual regions. The results of this study will aid in our understanding of the molecular epidemiology of PCV2.


Assuntos
Infecções por Circoviridae/epidemiologia , Infecções por Circoviridae/veterinária , Circovirus/classificação , Circovirus/genética , Variação Genética/genética , Genoma Viral/genética , Animais , China , Infecções por Circoviridae/virologia , Circovirus/isolamento & purificação , Coinfecção/virologia , Genótipo , Epidemiologia Molecular , Suínos
18.
Environ Pollut ; 247: 1134-1142, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30823342

RESUMO

Antibiotics are introduced into agricultural fields by the application of manure or biosolids, or via irrigation using reclaimed wastewater. Antibiotics can enter the terrestrial food chains through plant uptake, which forms an alternative pathway for human exposure to antibiotics. However, previous studies mainly focused on detecting residues of the parent antibiotics, while ignoring the identification of antibiotics transformation products in plants. Here, we evaluated the uptake and metabolism of clarithromycin (CLA) and sulfadiazine (SDZ) in lettuce under controlled hydroponic conditions. The antibiotics and their metabolites were identified by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QToF-MS/MS) and ultra-performance liquid chromatograph Micromass triple quadrupole mass spectrometry (UPLC-QqQ-MS/MS). The structure of CLA, SDZ and N-acetylated SDZ were confirmed with synthesized standards, verifying the reliability of the identification method. Eight metabolites of CLA and two metabolites of SDZ were detected in both the leaves and roots of lettuce. The metabolites of CLA included phases I and II transformation products, while only phase II metabolites of SDZ were observed in lettuce. The proportion of CLA metabolites was estimated to be greater than 70%, indicating that most of the CLA was metabolized in plant tissues. The proportion of SDZ metabolites was lower than 12% in the leaves and 10% in the roots. Some metabolites might have the ability to increase or acquire antibacterial activity. Therefore, in addition to the parent compounds, metabolites of antibiotics in edible vegetables are also worthy of study for risk assessment and to determine the consequences of long-term exposure.


Assuntos
Antibacterianos/metabolismo , Claritromicina/metabolismo , Alface/metabolismo , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Sulfadiazina/metabolismo , Verduras/metabolismo , Humanos
19.
Oncol Rep ; 41(4): 2351-2360, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30720133

RESUMO

The development of cervical cancer (CC) is a multi­gene, multi­step carcinogenic process that involves complex genetic and epigenetic mechanisms. SRY­related HMG­box gene 11 (SOX11) is a member of the SOX family of transcription factors with an emerging crucial role in the development of various tumor types. To elucidate the function of SOX11 in cervical carcinogenesis, the expression level of SOX11 during the development of human CC was analyzed by immunohistochemistry and western blot analysis. Additionally, the methylation status of the SOX11 was examined using bisulfite sequencing and methylation­specific polymerase chain reaction. The SOX11 expression and promoter methylation in human CC cell lines were also determined. The effect of SOX11 expression restoration after 5­aza­2'­deoxycytidine (5­Aza­dC) treatment on the CC cell proliferation ability was evaluated in CC cell lines. SOX11 was highly expressed in normal cervix (NC) and precancerous low­grade squamous intraepithelial lesions, but weakly expressed or virtually absent in precancerous high­grade squamous intraepithelial lesions and CC, which is consistent with the result of the western blot analysis. Hypermethylation of the SOX11 promoter was detected in CC, which was significantly higher than that in NC samples at each CpG site. The expression level of SOX11 in the CC cell lines was downregulated compared with the positive control, Tera­1human teratoma cell line. Upon 5­Aza­dC treatment, SOX11 expression was significantly upregulated in the CC cell lines at the mRNA and protein levels, and cell proliferation was inhibited. The results indicated that the downregulation of SOX11 in CC is due to the hypermethylation of the SOX11 promoter region. Thus, SOX11 methylation may have a role in the growth of CC cells and cervical carcinogenesis.


Assuntos
Regulação Neoplásica da Expressão Gênica , Fatores de Transcrição SOXC/genética , Proteínas Supressoras de Tumor/genética , Neoplasias do Colo do Útero/genética , Adulto , Azacitidina/farmacologia , Carcinogênese/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Colo do Útero/patologia , Ilhas de CpG/genética , Metilação de DNA/efeitos dos fármacos , Progressão da Doença , Regulação para Baixo/efeitos dos fármacos , Epigênese Genética , Feminino , Humanos , Pessoa de Meia-Idade , Regiões Promotoras Genéticas/genética , Fatores de Transcrição SOXC/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Regulação para Cima/efeitos dos fármacos , Neoplasias do Colo do Útero/patologia
20.
Mol Cell Probes ; 44: 44-50, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30735700

RESUMO

The development of a rapid, specific, and sensitive SYBR Green I-based duplex real-time quantitative PCR assay is described for the simultaneous detection of porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 3 (PCV3). The assay specifically detected PEDV and PCV3, with no fluorescence detected for other non-targeted pig pathogens. The assay showed a good linear relationship, and the limits of detection for this assay were 34.6 copies/µL and 61.2 copies/µL for PEDV and PCV3, respectively. The assay exhibited high repeatability and reproducibility, with intra-assay and inter-assay variation coefficients less than 2.0%. A clinical evaluation using intestinal tissue and fecal samples from piglets suffering from diarrhea at different pig farms in China revealed that the singular infection rates of PEDV and PCV3 were 43.94% (29/66) and 16.67% (11/66), respectively, while the co-infection rate of PCV3 with PEDV was 27.27% (18/66). The results indicate this assay is a rapid and reliable diagnostic tool for PEDV and PCV3 monitoring and surveillance in the field, and provides technical support for the quantitative detection of clinical samples infected or co-infected with PEDV and PCV3.


Assuntos
Circovirus/isolamento & purificação , Compostos Orgânicos/metabolismo , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Suínos/virologia , Animais , Circovirus/genética , Fluorescência , Desnaturação de Ácido Nucleico , Vírus da Diarreia Epidêmica Suína/genética , Padrões de Referência , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
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