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1.
Gene ; 764: 145097, 2021 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-32866589

RESUMO

Tomato (Lycopersicon esculentum Mill) is an important food plant that has been used as a model plant in genetic evolution and molecular biology research. The plant is originated from the tropics; thus, it is sensitive to cold. Its growth and development can be easily affected by cold stress. In this study, cold-regulated gene LeCOR413PM2 was cloned from tomato leaves and then used to generate two types of transgenic tomato plants: LeCOR413PM2-overexpressing transgenic plants and RNA-interference-expressing transgenic plants. The functions and expression of LeCOR413PM2 gene in response to cold stress were subsequently assessed. The results showed that LeCOR413PM2 localized in the plasma membrane. Expression of LeCOR413PM2 gene in the leaf of transgenic tomato plant was highest compared to that in other organs (i.e., root, stem, flower and fruit); it was elevated when plants were treated with cold stress. Overexpression of LeCOR413PM2 gene was found to not only reduce damage to cell membrane, accumulation of ROS, and photoinhibition of PSII, but also maintain high activity of antioxidant enzymes and content of osmotic regulators. The results also reveal that high activities of antioxidant enzymes were caused by the up-regulation of their gene expressions. This study demonstrates that the overexpression of LeCOR413PM2 could increase cold tolerance of transgenic tomato plants, while the suppressed expression of LeCOR413PM2 by RNA interference could increase the sensitivity of plants to cold.

2.
Rapid Commun Mass Spectrom ; : e8959, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-33001505

RESUMO

RATIONALE: Iris tectorum Maxim is a traditional medicinal herb that is commonly utilized for treating inflammatory conditions. The present study investigated the fragmentation patterns of isoflavone glycosides and their qualitative analysis. In addition, lipopolysaccharide (LPS)-induced RAW264.7 macrophages were used to evaluat the anti-inflammatory properties of I. tectorum Maxim samples collected at different time points during the year. METHOD: High-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (HPLC/Q-TOF-MS/MS) and HPLC-DAD were employed for the qualitative and quantitative analysis. The fragmentation patterns of these isoflavones were observed in negative electrospray ionization mode with collision-induced dissociation (CID). Their anti-inflammatory activity was assessed via nitric oxide (NO) production in LPS-treated RAW 264.7 macrophages. RESULTS: A total of 15 chemical components were observed and tentatively identified by HPLC/Q-TOF-MS. At low collision energy, the relative abundances of the aglycone radical anion Y0 - , [Y0 -H]-• , [Y0 -CH3 ]-• , and [Y0 -H-CH2 ]-• were used for the structural characterization of tectoridin and tectorigenin-4'-O-ß-D-glucoside. The radical ions [Y0 -CH3 ]-• and [Y0 -H-2CH3 ]-• were also employed to differentiate between iristectorin A and iristectorin B based upon their high energy CID spectra. Levels of 9.02 mg/g of tectoridin and 1.04 mg/g of tectorigenin were found in samples collected in June, which exhibited 69.7% NO inhibitory activity. CONCLUSION: The characteristic fragmentation patterns enabled us to reliably identify isoflavone glycosides. The results of the quantitative determination and NO inhibitory activity offer insight into the optimal I. tectorum Maxim harvesting time.

3.
Artigo em Inglês | MEDLINE | ID: mdl-33004280

RESUMO

OBJECTIVE: Thoracic aortic dissection (TAD) is associated with matrix changes, biochemical changes, and inflammatory markers like interleukin-1 beta (IL-1ß). However, the exact mechanism remains unknown. This study aimed to investigate the role of IL-1ß, matrix metalloproteinase (MMP)-2, MMP-9, smooth muscle cell apoptosis, and elastic fibre fracture in the development of TAD in a rat model. METHODS: The TAD rat model was induced by ß-aminopropionitrile (BAPN). TAD was investigated in 112 male Sprague-Dawley rats, which were equally divided into four groups of 28 rats (Control, BAPN, BAPN + IL-1ß, and BAPN + IL-1ß antibody). Systolic blood pressure, survival, and the development of TAD were measured after six weeks. Expression of IL-1ß, MMP-2, and MMP-9 was measured by Western blot. Apoptosis, aortic elastin concentration, and biomechanical characteristics were measured by the TdT mediated dUTP nick end labelling assay, Victoria blue staining, and in vitro testing. RESULTS: During six weeks, the mortality was 0% (0/28) in the control group, 53.6% (15/28) in the BAPN group (p < .001 compared with the control group), 75.0% (21/28) in the BAPN + IL-1ß group (p = .007 compared with the BAPN group), and 35.7% (10/28) in the BAPN + IL-1ß antibody group (p = .023 compared with BAPN group and p < .001 compared with the BAPN + IL-1ß group). IL-1ß treatment deteriorates BAPN induced mortality and aneurysm expansion, which were attenuated by anti-IL-1ß treatment. In BAPN + IL-1ß group, stress and strain parameters were decreased by 13.5%-53.5% and elastin content was decreased by 14%, and IL-1ß, MMP-2, and MMP-9 were expressed higher by 117%, 108%, and 75% when compared with the rats in the BAPN group. Contrarily, in the BAPN + IL-1ß antibody group, the above changes could be completely (strain, elastin content, and expression of MMP-2) or partly (elasticity modulus, stress, and expression of MMP-9) blocked by anti-IL-1ß treatment. CONCLUSION: IL-1ß plays a critical role in TAD formation by altering the expression of MMP-2 and MMP-9, degrading the aortic wall matrix, causing elastic fibre rupture, and changing the stress or strain of the aortic wall. Anti-IL-1ß reduces the later effects and could be one of the molecular targets for prognosis and drug treatment of TAD in the future.

4.
Oxid Med Cell Longev ; 2020: 7582980, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33005288

RESUMO

Mitochondrial damage in airway epithelial cells plays an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). Sirtuin 3 (Sirt3) is a mitochondrial deacetylase regulating mitochondrial function, but its role in the pathogenesis of COPD is still unknown. The aim of the present study was to investigate the effect of Sirt3 on airway epithelial mitochondria in cigarette smoke-induced COPD. Our present study has shown serious airway inflammation, alveolar space enlargement, and mitochondrial damage of the airway epithelium in COPD rats. Compared to the control rats, Sirt3 protein expression was significantly decreased in the airway epithelium and lung tissue homogenate from COPD rats. In airway epithelial cells (BEAS-2B), cigarette smoke extract (CSE) treatment significantly decreased mRNA and protein expression of Sirt3 and manganese superoxide dismutase (MnSOD), as well as MnSOD activity in a concentration and time-dependent manner. Sirt3 siRNA further significantly intensified the decreases in MnSOD expression and activity and aggravated mitochondrial oxidative stress and cell injury when airway epithelial cells were treated with 7.5% CSE. In contrast, Sirt3 overexpression significantly prevented the decrease of MnSOD expression and activity and improved mitochondrial oxidative stress and cell injury in CSE-treated airway epithelial cells. These data suggest that Sirt3 inhibits airway epithelial mitochondrial oxidative stress possibly through the regulation of MnSOD, thereby contributing to the pathogenesis of COPD.

5.
Biochim Biophys Acta Rev Cancer ; : 188441, 2020 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-33007432

RESUMO

A couple of molecular-targeting medications, such as Lenvatinib, are available for the treatment of hepatocellular carcinoma (HCC) in addition to Sorafenib in an advanced stage. Approval for the use of immune check-point inhibitors, such as Nivolumab and Pembrolizumab has shifted the paradigm of current HCC treatment, and the monotherapy or in combination with Lenvatinib or Sorafenib has significantly extended overall survival or progression-free survival in a large portion of patients. A combination of programmed cell death ligand-1 (PD-L1) inhibitor Atezolizumab with a vascular endothelial growth factor (VEGF) inhibitor, Bevacizumab, has recently achieved promising outcome in unresectable HCC patients. Other immunotherapy, such as chimeric antigen receptor T (CAR-T) cell therapy has achieved an evolutional success in hematologic malignancies, and has extended its use in deadly solid tumors, such as HCC. Although there exist various barriers, novel approaches are developed to move potential adoptive T cell therapy strategies, including cytokine-induced killer (CIK) cells, tumor-infiltrating lymphocytes (TIL), T cell receptor (TCR) T cells, CAR-T cells, to clinical application.

6.
Oncogene ; 2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-33009488

RESUMO

Neuroendocrine prostate cancer (NEPC) is an aggressive variant of prostate cancer that either develops de novo or arises from prostate adenocarcinoma as a result of treatment resistance. Although the prostate basal cells have been shown to directly generate tumor cells with neuroendocrine features when transduced with oncogenic signaling, the identity of the cell-of-origin for de novo NEPC remains unclear. We show that the TACSTD2high human prostate luminal epithelia cells highly express SOX2 and are relatively enriched in the transition zone prostate. Both TACSTD2high and TACSTD2low luminal cells transduced by constitutively activated AKT1 (caAKT1), and c-Myc can form organoids containing versatile clinically relevant tumor cell lineages with regard to the expression of AR and the neuroendocrine cell markers Synaptophysin and Chromogranin A. Tumor organoid cells derived from the TACSTD2high luminal cells are more predisposed to neuroendocrine differentiation along passaging and are relatively more castration-resistant. Knocking down TACSTD2 and SOX2 both attenuate neuroendocrine differentiation of tumor organoid cells. This study demonstrates de novo neuroendocrine differentiation of the human prostate luminal epithelial cells induced by caAKT1 and c-Myc and reveals an impact of cellular status on initiation of lineage plasticity.

7.
J Hum Genet ; 2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-33009504

RESUMO

Polygenic risk scores (PRS) estimate the genetic risk of an individual for a complex disease based on many genetic variants across the whole genome. In this study, we compared a series of computational models for estimation of breast cancer PRS. A deep neural network (DNN) was found to outperform alternative machine learning techniques and established statistical algorithms, including BLUP, BayesA, and LDpred. In the test cohort with 50% prevalence, the Area Under the receiver operating characteristic Curve (AUC) were 67.4% for DNN, 64.2% for BLUP, 64.5% for BayesA, and 62.4% for LDpred. BLUP, BayesA, and LPpred all generated PRS that followed a normal distribution in the case population. However, the PRS generated by DNN in the case population followed a bimodal distribution composed of two normal distributions with distinctly different means. This suggests that DNN was able to separate the case population into a high-genetic-risk case subpopulation with an average PRS significantly higher than the control population and a normal-genetic-risk case subpopulation with an average PRS similar to the control population. This allowed DNN to achieve 18.8% recall at 90% precision in the test cohort with 50% prevalence, which can be extrapolated to 65.4% recall at 20% precision in a general population with 12% prevalence. Interpretation of the DNN model identified salient variants that were assigned insignificant p values by association studies, but were important for DNN prediction. These variants may be associated with the phenotype through nonlinear relationships.

8.
Infect Dis Poverty ; 9(1): 136, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-33004080

RESUMO

BACKGROUND: The People's Republic of China (P. R. China) has made significant progress on schistosomiasis control. Among the 12 provincial-level administrative divisions (PLADs) with schistosomiasis endemic in P. R. China, Guangdong, Shanghai, Fujian, Guangxi and Zhejiang PLADs (following as five PLADs) had successively eliminated schistosomiasis during 1985-1995. However, consolidation of the schistosomiasis elimination in these five PLADs remains challenging. In the current study, we sought to understand the epidemic situation in these post-elimination areas and their surveillance capabilities on schistosomiasis. METHODS: Annual data reflecting the interventions and surveillance on human beings, cattle and snails based on county level from 2005 to 2016 were collected through the national schistosomiasis reporting system and the data were analyzed to understand the epidemic status of schistosomiasis in the five PLADs. A standardized score sheet was designed to assess the surveillance capacity for schistosomiasis of selected disease control agencies in five PLADs and ten counties. Assessment on surveillance capacity including schistosomiasis diagnostic skills, identification of snails' living and infection status and knowledge about schistosomiasis and its control were made. Descriptive analysis was used to analyze the epidemic status and evaluation results on surveillance capacities. RESULTS: The assessments showed that no local cases in humans and cattle or infected snail were found in these five PLADs since 2005. However, from 2005 to 2016, a total of 221 imported cases were detected in Zhejiang, Shanghai and Fujian, and 11.98 hm2 of new snail habitats were found in Zhejiang, Shanghai and Guangxi. In addition, snail infestation reoccurred in 247.55 hm2 of former snail habitats since 2011. For the surveillance capacity assessment, the accuracy rate of IHA and MHT were 100 and 89.3%, respectively. All participants could judge the living status of snails accurately and 98.1% on the infection status of snails. The accuracy rate of the questionnaire survey was 98.0%. CONCLUSIONS: Elimination of schistosomiasis was consolidated successfully in five PLADs of P. R. China due to effective and strong post-elimination surveillance. Comprehensive consolidation strategies should be focused on the elimination of residual snails and the prevention of imported infection sources to consolidate the achievements of schistosomiasis control.

9.
Eur J Appl Physiol ; 2020 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-33025229

RESUMO

PURPOSE: We examined the possibility that wearing a below-knee compression garment (CG) reduces fatigue-induced strength loss and joint position sense (JPS) errors in healthy adults. METHODS: Subjects (n = 24, age = 25.5 ± 4 years) were allocated to either one of the treatment groups that performed 100 maximal isokinetic eccentric contractions at 30°-1 with the right-dominant knee extensors: (1) with (EXPCG) or (2) without CG (EXP) or to (3) a control group (CONCG: CG, no exercise). Changes in JPS errors, and maximal voluntary isometric contraction (MVIC) torque were measured immediately post-, 24 h post-, and 1 week post-intervention in each leg. All testing was done without the CG. RESULTS: CG afforded no protection against JPS errors. Mixed analysis of variance (ANOVA) revealed that absolute JPS errors increased post-intervention in EXPCG and EXP not only in the right-exercised (52%, p = 0.013; 57%, p = 0.007, respectively) but also in the left non-exercised (55%, p = 0.001; 58%, p = 0.040, respectively) leg. Subjects tended to underestimate the target position more in the flexed vs. extended knee positions (75-61°: - 4.6 ± 3.6°, 60-50°: - 4.2 ± 4.3°, 50-25°: - 2.9 ± 4.2°), irrespective of group and time. Moreover, MVIC decreased in EXP but not in EXPCG and CONCG at immediately post-intervention (p = 0.026, d = 0.52) and 24 h post-intervention (p = 0.013, d = 0.45) compared to baseline. CONCLUSION: Altogether, a below-knee CG reduced fatigue-induced strength loss at 80° knee joint position but not JPS errors in healthy younger adults.

10.
Artigo em Inglês | MEDLINE | ID: mdl-33013682

RESUMO

Background: Papillary thyroid carcinoma (PTC) patients with anterior extrathyroidal extension (ETE) involving the strap muscle have a relatively better prognosis than those with posterior gross ETE involving the recurrent laryngeal nerve. Whether prophylactic central-compartment lymph node dissection (CLND) should be performed in PTCs with only strap muscle invasion (SMI) is still unclear. Methods: A retrospective cohort study was conducted in clinical N0 (cN0) PTC patients with SMI who underwent thyroid surgery from 2009 to 2017. A total of 152 patients were included, and predictive factors of central-compartment lymph node metastasis (CLNM) were determined. Results: Among the 281 PTCs patients with SMI, 152 (51.1%) did not clinically present with lymph node metastasis. Microscopic CLNM was identified in 77 (50.7%) cN0 PTC patients with SMI. According to the univariate and multivariate analyses, male patients and those aged <40 years were more likely to be diagnosed with CLNM than female patients and those aged >40 years (odds ratio [OR] = 6.22 [95% confidence interval (CI), 1.43-27.10], p = 0.02 vs. OR = 9.94 [95% CI, 2.79-35.44], p = 0.00). The CLNM positive rate of male patients aged <40 years was 87.5%, while that for female patients aged ≥55 years was 23.8%. However, risk factors associated with large-volume CLNM were not identified because of the small number of patients. Conclusions: Taken together, nearly half of PTC patients with SMI did not clinically present with lymph node metastasis. Male sex and patients aged <40 years were identified as the predictive factors of CLNM in cN0 PTCs with SMI. Hence, the results of this single-center study raise the possibility that prophylactic CLND may be more often considered for younger male PTC patients with SMI.

11.
Artigo em Inglês | MEDLINE | ID: mdl-33013688

RESUMO

Objective: We aimed to assess the role of adipose tissue distribution in cardiometabolic risk (in particular insulin sensitivity) in a population of children and adolescents with obesity. Methods: In this cross-sectional study, participants were 479 children and adolescents with obesity (322 boys and 157 girls) aged 3 to 18 years attending the Children's Hospital at Zhejiang University School of Medicine (Hangzhou, China). Clinical assessments included anthropometry, body composition (DXA scans), carotid artery ultrasounds, and OGTT. Insulin sensitivity was assessed using the Matsuda index. Participants were stratified into groups by sex and pubertal stage. Key predictors were DXA-derived android-to-gynoid-fat ratio (A/G) and total body fat percentage (TBF%). Results: Irrespective of sex and pubertal stage, there was a strong association between increasing A/G (i.e., greater abdominal adiposity) and lower insulin sensitivity. In multivariable models, every 0.1 increase in A/G was associated with a reduction in insulin sensitivity in prepubertal boys [-29% (95% CI -36%, -20%); p < 0.0001], pubertal boys [-13% (95% CI -21%, -6%); p = 0.001], and pubertal girls [-16% (95% CI -24%, -6%); p = 0.002]. In contrast, TBF% was not associated with insulin sensitivity when A/G was adjusted for, irrespective of pubertal stage or sex. In addition, every 0.1 increase in A/G was associated with increased likelihood of dyslipidemia in prepubertal boys [adjusted odds ratio (aOR) 1.62 (95% CI 1.05, 2.49)], impaired glucose tolerance in pubertal boys [aOR 1.64 (95% CI 1.07, 2.51)] and pubertal girls [aOR 1.81 (95% CI 1.10, 2.98)], and odds of NAFLD in both prepubertal [aOR 2.57 (95% CI 1.56, 4.21)] and pubertal [aOR 1.69 (95% CI 1.18, 2.40)] boys. In contrast, higher TBF% was only associated with higher fasting insulin and ALT in pubertal boys, being also predictive of NAFLD in this group [aOR 1.15 per percentage point (95% CI 1.06, 1.26)], but was not associated with the likelihood of other cardiometabolic outcomes assessed in any group. Conclusions: A/G is a much stronger independent predictor of cardiometabolic risk factors in children and adolescents with obesity in China, particularly glucose metabolism.

12.
Front Immunol ; 11: 1717, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33013823

RESUMO

Vascular endothelial injury caused by post-hemorrhagic shock mesenteric lymph (PHSML) return is an important manifestation during refractory hemorrhagic shock. Using human umbilical vein endothelial cells (HUVECs) and transcriptome analysis, this study sought to investigate the molecular mechanism underlying the adverse effect of PHSML on vascular endothelium. Post-hemorrhagic shock mesenteric lymph was collected from male rats after they underwent hemorrhagic shock and following resuscitation, while normal mesenteric lymph (NML) was harvested from sham rats. Human umbilical vein endothelial cells were incubated with the culture medium containing either 10% phosphate buffered saline (Control), NML, or PHSML for 3 h, and then were harvested for RNA sequencing. In comparison with NML treated cells, 37 genes were differentially expressed in PHSML-treated HUVECs, including 32 upregulated genes and five downregulated genes. These differentially expressed genes were mainly enriched in inflammatory pathways, including signaling pathways for activation of the NOD-like receptors, NF-κB, and TNF. Furthermore, we found that C-C motif chemokine ligand 2 (CCL2) was increased significantly after PHSML treatment, and Bindarit, a CCL2 production inhibitor, attenuated the damage of HUVECs induced by PHSML. The results provide molecular evidence on vascular endothelium damage caused by PHSML. C-C motif chemokine ligand 2 might represent a new target for reducing vascular injury after severe hemorrhagic shock.

13.
Artigo em Inglês | MEDLINE | ID: mdl-33019979

RESUMO

Hyperglycemia promotes podocyte apoptosis and plays an important role in the pathogenesis of diabetic nephropathy (DN). Calcium/calcineurin (CaN) signaling is critical for podocyte apoptosis. Therefore, it is essential to elucidate the mechanisms underlying the regulation of CaN signaling. Recent studies reported that histone deacetylase 4 (HDAC4) is involved in podocyte apoptosis in DN. The aim of this study was to determine whether HDAC4 mediates the regulation of CaN and to elucidate the function of HDAC4 in high glucose (HG)-induced podocyte apoptosis. First, we identified the expression of HDAC4 was upregulated in podocytes of patients with DN. In vitro, the results also indicate that the mRNA and protein expression levels of HDAC4 were increased in HG-cultured podocytes. Silencing and overexpression of HDAC4 markedly decreased and increased CaN expression, respectively. Meanwhile, HG-induced podocyte apoptosis was abrogated by HDAC4-knockdown with subsequent decreased Bax expression and increased Bcl-2 expression. In contrast, overexpression of HDAC4 increased podocyte apoptosis and Bax expression, as well as decreased Bcl-2 expression. In addition, podocyte apoptosis induced by HDAC4 overexpression was effectively rescued by FK506, a pharmacological inhibitor of CaN, which was accompanied by decreased Bax and increased Bcl-2 expression. As a novel finding, HG-induced podocyte apoptosis is mediated by the HDAC4/CaN signaling pathway, which presents a promising target for therapeutic intervention in DN.

14.
Artigo em Inglês | MEDLINE | ID: mdl-33011298

RESUMO

BACKGROUND: Hospitalization due to acute asthma exacerbation (AE) is a highly detrimental situation requiring critical management to prevent further deterioration, including mechanical ventilation, intensive care unit (ICU) admission, and death. However, patients hospitalized for AEs are highly heterogeneous and remain largely unexplored. OBJECTIVE: Identify clinical and inflammatory phenotypes of AE requiring hospitalization associated with in-hospital outcomes. METHODS: We performed a hierarchical cluster analysis of 825 consecutively recruited patients hospitalized for AEs. Logistic regressions were conducted to quantify the independent associations of the identified phenotypes with in-hospital outcomes. Decision tree analysis was developed to predict cluster assignment. RESULTS: We identified three clusters of patients, which had significantly different characteristics associated with in-hospital adverse outcomes. Cluster 1 (n=526, 63.8%) was a late-onset phenotype, cluster 2 (n=97, 11.8%) was an early-onset phenotype, and cluster 3 (n=202, 24.5%) was a phenotype with fewer eosinophils and more comorbidities. Clusters 2 and 3 had an elevated risk of death (RRadj 18.10 and 19.17, respectively) and mechanical ventilation (RRadj 2.56 and 5.71, respectively) than did cluster 1. Individuals in cluster 3 had an extended length of hospital stay (11 days), increased hospitalization direct costs (13,481.57 Chinese Yuan[CNY]), and a higher risk of ICU admission (RRadj 2.14) than individuals in clusters 1 and 2. The decision tree assigned 90.8% of the participants correctly. CONCLUSIONS: We identified three phenotypes with differential clinical and inflammatory characteristics associated with in-hospital adverse outcomes. These new phenotypes might have important and clinically relevant implications for the management of patients hospitalized for AEs.

15.
Biochem Genet ; 2020 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-33011918

RESUMO

Aspidopterys obcordata var. obcordata, a medicinal plant endemic to China, is a narrowly distributed species and wild resources are extremely limited. To evaluate the genetic variability and degree of genetic divergence of A. obcordata var. obcordata, and to make rational scientific decisions on its harvest and germplasm conservation, we collected 122 samples from across nearly all of its distribution area and studied genetic diversity using inter-simple sequence repeats (ISSRs), sequence-related amplified polymorphisms (SRAPs), and a method combining the two techniques. The results revealed the high genetic diversity of A. obcordata var. obcordata, mainly due to its intra-population diversity, and the top two populations with the highest levels of intra-population diversity were ML and DH, individuals of which can serve as excellent germplasm candidates during the processing of germplasm screening and conservation. In general, the combining method was prior to the ISSR analyses and SRAP analyses results, except for a slight difference in the genetic structure of individual populations. Therefore, we suggest that a combination analysis of the two marker methods is ideal for evaluating the genetic diversity and genetic relationships of A. obcordata var. obcordata.

16.
Artigo em Inglês | MEDLINE | ID: mdl-33012221

RESUMO

Identification of matrix bound nanovesicles (MBV) as ubiquitous components of the extracellular matrix (ECM) raises questions regarding their biologic functions and their potential theranostic application. Unlike liquid-phase extracellular vesicles (e.g., exosomes), MBV are tightly bound to the ECM which makes their isolation and harvesting more challenging. The indiscriminate use of different methods to harvest MBV can alter or disrupt their structural and/or functional integrity. The objective of the present study was to compare the effect of various MBV harvesting methods upon yield, purity, and biologic activity. Combinations of four methods to solubilize the ECM (collagenase, liberase, or proteinase K, and non-enzymatic elution with potassium chloride) and four isolation methods (ultracentrifugation, ultrafiltration, density barrier, and size exclusion chromatography) were used to isolate MBV from urinary bladder derived ECM. All combinations of solubilization and isolation methods allowed for the harvesting of MBV, however distinct differences were noted. The highest yield, purity, cellular uptake, and biologic activity were seen with MBV isolated by a combination of liberase or collagenase followed by size exclusion chromatography. The combination of proteinase K and ultrafiltration was shown to have detrimental effects on bioactivity. The results show the importance of selecting appropriate MBV harvesting methods for the characterization and evaluation of MBV and for analysis of their potential theranostic application.

17.
Artigo em Inglês | MEDLINE | ID: mdl-33016781

RESUMO

Background: Previous studies have shown puerarin to be a potential therapeutic drug for treatment of bladder cancer. But the role and possible molecular mechanism of puerarin remain unknown. Methods: Cell viability, apoptosis, migration, and invasion were assessed by Cell Counting Kit-8 (CCK-8), flow cytometry, and transwell assays, respectively. Western blot was used to measure the levels of all protein. Glucose consumption and lactate production were detected using a glucose and lactate assay kit. Circular RNA_0020394 (circ_0020394), microRNA-328-3p (miR-328-3p), and nuclear receptor binding protein 1 (NRBP1) levels were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The interaction between miRNA and circRNA or mRNA was confirmed using dual-luciferase reporter assay. In vivo experiments were performed to examine the effect of puerarin on tumor growth. Results: Puerarin suppressed cell viability, migration, invasion, and glycolysis, and induced apoptosis in bladder cancer. circ_0020394 was downregulated in puerarin-treated bladder cancer cells, and circ_0020394 overexpression attenuated the inhibitory effect of puerarin on cell progression. Moreover, circ_0020394 could bind to miR-328-3p, and miR-328-3p directly targeted NRBP1. Functionally, miR-328-3p could reverse the promotion effect of circ_0020394 overexpression on the progression of puerarin-treated cells, and silencing NRBP1 counteracted the effects of anti-miR-328-3p on puerarin-treated cells. Mechanically, circ_0020394 could increase NRBP1 expression by acting as miR-328-3p sponge in puerarin-treated bladder cancer cells. Besides, puerarin inhibited tumorigenesis in vivo by increasing miR-328-3p and decreasing the levels of circ_0020394 and NRBP1. Conclusions: Puerarin impedes cell viability, migration, invasion, and glycolysis, and promoted apoptosis in bladder cancer by regulating circ_0020394/miR-328-3p/NRBP1 axis.

18.
J Biomol Struct Dyn ; : 1-10, 2020 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-33016857

RESUMO

Wee1 kinase and Checkpoint kinase 1 (Chk1) kinase, which are well known to be involved in cancer, are promising targets for cancer therapy. Most of developed Wee1 inhibitors can inhibit activity of Chk1 kinase to different degrees as well. The poor selectivity brought side effects and selective inhibitor is needed. However, the selective mechanisms of Wee1 versus Chk1 are not clear. Therefore, the design of selective Wee1 and Chk1 inhibitors would provide a meaningful starting for the development of anticancer drugs with optimal efficacy. In this study, Wee1 inhibitors with different selectivity over Chk1 were chosen to analyze the selectivity mechanism by means of molecular docking, molecular dynamics simulations and binding free energy calculations. Two key residues of Wee1 kinase and two critical residues of Chk1 were mutated to detect their effect on ligand binding into protein. The results indicated that these residues play a pivotal role in the binding interactions of ligands to receptors through hydrogen bond and hydrophobic interaction with inhibitors. This may provide a better understanding of the selective mechanism of Wee1 and Chk1. It would be beneficial to the discovery and optimization of selective Wee1 and Chk1 inhibitors. Communicated by Ramaswamy H. Sarma.

19.
Technol Health Care ; 2020 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-33016898

RESUMO

BACKGROUND: Minimally invasive total knee arthroplasty (TKA) has been actively advertised by the orthopedic industry. The purpose of this network meta-analysis was to comprehensively compare the effectiveness of four minimally invasive surgery (MIS) approaches and the medial parapatellar (MPP) approach to improve the American Knee Society Score (KSS) in primary TKA. MATERIALS AND METHODS: Studies were comprehensively searched on PubMed, Embase, Cochrane Library (CENTRAL), Web of Science and Science Direct up to June 2018 with a major focus on the outcome of KSS. Risk of bias was assessed using the Cochrane risk of bias tool. Quality assessment was performed using the GRADE system. Both pair-wise and network meta-analyses are calculated to comprehensively compare the effectiveness of four MIS and TKA approaches. RESULTS: Eleven trials with 1025 knees undergoing TKA were included. Our analysis showed that both MPP and MIS approaches provided improvement in terms of short-term (four-eight weeks) total, objective and functional KSS score. The network-meta analysis revealed that MIS approaches showed a trend towards superior KSS improvement over standard MPP approaches. However, statistical advantages were only observed in the mini-MPP group for functional KSS scores when compared with conventional MPP and quadriceps-sparing (QS) group. CONCLUSIONS: Evidence shows that MIS TKA approaches are effective alternatives to MPP approaches. However, orthopedic surgeons should be cautiously optimistic about minimally invasive TKA in terms of KSS improvement.

20.
Plant Genome ; 13(1): e20001, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-33016624

RESUMO

African wild rice Oryza longistaminata, one of the eight AA- genome species in the genus Oryza, possesses highly valued traits, such as the rhizomatousness for perennial rice breeding, strong tolerance to biotic and abiotic stresses, and high biomass production on poor soils. To obtain the high-quality reference genome for O. longistaminata we employed a hybrid assembly approach through incorporating Illumina and PacBio sequencing datasets. The final genome assembly comprised only 107 scaffolds and was approximately ∼363.5 Mb, representing ∼92.7% of the estimated African wild rice genome (∼392 Mb). The N50 lengths of the assembled contigs and scaffolds were ∼46.49 Kb and ∼6.83 Mb, indicating ∼3.72-fold and ∼18.8-fold improvement in length compared to the earlier released assembly (∼12.5 Kb and 364 Kb, respectively). Aided with Hi-C data and syntenic relationship with O. sativa, these assembled scaffolds were anchored into 12 pseudo-chromosomes. Genome annotation and comparative genomic analysis reveal that lineage-specific expansion of gene families that respond to biotic- and abiotic stresses are of great potential for mining novel alleles to overcome major diseases and abiotic adaptation in rice breeding programs. This reference genome of African wild rice will greatly enlarge the existing database of rice genome resources and unquestionably form a solid base to understand genomic basis underlying highly valued phenotypic traits and search for novel gene sources in O. longistaminata for the future rice breeding programs.


Assuntos
Oryza , Genoma , Genômica , Oryza/genética , Análise de Sequência de DNA
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