Oral polysaccharide-coated liposome-modified double-layered nanoparticles containing anthocyanins: preparation, characterization, biocompatibility and evaluation of lipid-lowering activity in vitro.

Anthocyanins (ACNs) systems encapsulated in nanomaterials have received widespread attention and rapid development due to its good delivery potential. Here, the favorable benefits of four natural polysaccharide food additives coated ACNs-liposome nanoparticles (ACNs-Lipo NPs) on the stability and possible lipid-lowering effects of ACNs are discussed in this work. The polysaccharides were coupled to the ACNs-Lipo NPs and self-assembled to create ACNs-Lipo@polysaccharide NPs. The impact of various polysaccharides on the physical, chemical, and stability characteristics of NPs was examined. We found that the NPs prepared with gum arabic (GA) had the best stability. FT-IR and XRD analysis revealed electrostatic adsorption and hydrogen binding forces between the components, as well as an amorphous structure. A series of tests in vitro confirmed the excellent stability, bioavailability, antioxidant activity, and biocompatibility of NPs. Finally, cellular antioxidant activity (CAA) and oleic acid (OA)-induced lipid deposition cell models revealed that ACNs-Lipo@GA might be more readily absorbed by cells, resulting in improved antioxidant activity and lipid-lowering impact, with possible targeted delivery qualities and lipid-lowering effect.

A comprehensive diagnostic scheme of morphological combined molecular methylation under bronchoscopy.

Methylated SHOX2 and RASSF1A genes are potential biomarkers for lung cancer diagnosis. Therefore, we explored the role of methylation detection combined with morphological bronchoscopic evaluation for lung cancer diagnosis. Bronchoscopy, methylation outcome, and pathological data were collected from 585 patients with lung cancer and 101 controls. The methylation status of the SHOX2 and RASSF1A genes were detected using real-time polymerase chain reaction quantification. Further, the sensitivity and area under the receiver operating characteristic curve of the three methods were analyzed. Among 686 patients, 57.1% had new lesions detected through bronchoscopy and 93.1% of these patients were diagnosed with malignant tumors. Besides, 42.9% of patients had no visible changes under bronchoscopy but there were still 74.8% of them diagnosed with malignant tumors. Bronchoscopy revealed that lung adenocarcinoma, lung squamous cell carcinoma, and small cell lung cancer mainly occurred in the upper and middle lobes. The sensitivity and specificity of methylation detection were 72.8% and 87.1% (vs. cytology 10.4% & 100%), respectively. Therefore, methylated SHOX2 and RASSF1A genes may be promising tumor markers in lung cancer diagnosis. Methylation detection can be an excellent supplementary tool for cytological diagnosis and, combined with bronchoscopy, could form a more effective diagnostic process.

A Bayesian model to identify multiple expression patterns with simultaneous FDR control for a multi-factor RNA-seq experiment.

It is often of research interest to identify genes that satisfy a particular expression pattern across different conditions such as tissues, genotypes, etc. One common practice is to perform differential expression analysis for each condition separately and then take the intersection of differentially expressed (DE) genes or non-DE genes under each condition to obtain genes that satisfy a particular pattern. Such a method can lead to many false positives, especially when the desired gene expression pattern involves equivalent expression under one condition. In this paper, we apply a Bayesian partition model to identify genes of all desired patterns while simultaneously controlling their false discovery rates (FDRs). Our simulation studies show that the common practice fails to control group specific FDRs for patterns involving equivalent expression while the proposed Bayesian method simultaneously controls group specific FDRs at all settings studied. In addition, the proposed method is more powerful when the FDR of the common practice is under control for identifying patterns only involving DE genes. Our simulation studies also show that it is an inherently more challenging problem to identify patterns involving equivalent expression than patterns only involving differential expression. Therefore, larger sample sizes are required to obtain the same target power to identify the former types of patterns than the latter types of patterns.

Structural characterization and hepatoprotective activity of exopolysaccharide from Bacillus velezensis SN-1.

BACKGROUND: Exopolysaccharide biopolymers produced by microorganisms are crucial to the environment. They contribute to areas such as the health and bionanotechnology sectors, food and cosmetic industries as gelling agents, and environmental sector as flocculants owing to their biodegradability and non-toxic nature. The current study aimed to isolate the fraction of released exopolysaccharide (rEPS) by Bacillus velezensis SN-1 from Chinese Da-Jiang. RESULTS: The weighted average molecular weight of the major isolated component, rEPS-2, was 202 kDa, and its monosaccharide composition included mannose, glucose, and galactose at a molar ratio of 0.38:0.30:0.32. Further, the rEPS-2 was characterized using methylation analysis and one-dimensional/two-dimensional nuclear magnetic resonance (1D/2D NMR) spectroscopy. In vivo hepatoprotective effects indicated that rEPS-2 could alleviate carbon tetrachloride (CCl4 )-induced liver injury in mice by lowering the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and the malondialdehyde (MDA) levels. Furthermore, rEPS-2 can increase the expression of antioxidant genes HO-1, GCLC and NQO1 in the Nrf2/ARE signaling pathway, thereby increasing the activity of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and reduced catalase (CAT) in liver cells. Furthermore, the rEPS-2 can be used and modulate the gut microbiota of mice with liver injury caused by CCl4 . CONCLUSIONS: These results suggest that rEPS-2 has promising potential to serve as hepatoprotective agents. © 2022 Society of Chemical Industry.

Antioxidant potential and in vitro inhibition of starch digestion of flavonoids from Crataegus pinnatifida.

Hawthorn flavonoids were extracted by enzymolysis associated ultrasonic procedure. Thirteen flavonoids were identified by HPLC/ESI-QTOF/MS, and the major components were procyanidin C1, rutin-rhamnoside, vitexin-rhamnoside, and catechin. Hawthorn flavonoids exhibited strong free radical scavenging activities against DPPH, ABTS, and hydroxyl radicals. Total and intercellular antioxidant experiments revealed that the free hydro-PSC value was 295.32 ± 12.20 µmol of VCE/g DW, and the free cellular antioxidant activity (CAA) values were 168.60 ± 4.87 µmol of QE/g DW in the no PBS wash protocol and 49.53 ± 1.75 µmol of QE/g DW in the PBS wash protocol. In addition, hawthorn flavonoids exhibited higher α-amylase and α-glucosidase inhibitory activities. The wheat starch digestibility was also reduced by hawthorn flavonoids as well. The results indicated that enzymolysis associated ultrasonic extraction was advisable for extracting flavonoids from hawthorn, and hawthorn flavonoids might be recommended as a potential food supplement with hypoglycemic activities.

Conversion of condensed tannin from chokeberry to cyanidin: Evaluation of antioxidant activity and gut microbiota regulation.

Chokeberries contain a large amount condensed tannin. In this study, hot acid-alcohol treatment was used to convert the condensed tannin into cyanidin, so as to give them higher antioxidant activity and improve the function of gut microbiota. The total cyanidins yield was taken as the index. The effects of various factors on the total cyanidins yield were analysed by the single factor experiment and factor design experiment. In addition, the response surface methodology was used to obtain the optimal conversion conditions. The cyanidin composition and cellular antioxidant activity of products was detected. Through simulated digestion and fermentation in vitro, improvement of gut microbiota was evaluated. The results show that the product obtained by the optimal conversion treatment conditions, which could increase total cyanidins yield 1.50 folds to 28.88 mg/g. The total antioxidant activity and cellular antioxidant activity increased by 1.85 folds and 1.56 folds, reaching 3905.48 (µmol Vit C equiv./100 g FW) and 2329.31 (µmol QE/100 g FW), respectively. Conversion treatment could change the some negative effect of chokeberry on gut microbiota diversity into the positive effect.

The effect of aerobic exercise on the lipophagy of adipose tissue in obese male mice.

This article explores the obesity state and the changes in the level of lipophagy in adipose tissue after exercise to lose weight, so as to provide direction and basis for theoretical research on obesity prevention and control. We established a high-fat diet model of obese mice, and applied exercise intervention and intraperitoneal injection of chloroquine to inhibit autophagy. Long-term high-fat diet can cause obesity in mice, and the process of lipophagy is inhibited, which may be one of the reasons for fat accumulation. Eight weeks of aerobic exercise can effectively reduce the weight of obese mice and promote lipolysis; this process is mainly completed by lipase decomposition, in addition to require the participation of the lipophagy process.

Power priors with entropy balancing weights in data augmentation of partially controlled randomized trials.

In pediatric or orphan diseases, there are many instances where it is unfeasible to conduct randomized and controlled clinical trials. This is due in part to the difficulty of enrolling a sufficient number of patients over a reasonable time period to meet adequate statistical power to demonstrate the treatment efficacy. One solution to reduce the sample size or expedite the trial timeline is to complement the current trial with real-world data. To this end, several propensity score-based methods have been developed to create defined groups of patients that are controlled for confounding based on a set of measured covariates at baseline. However, balance checking on the measured covariates and tweaks to the propensity score models is usually inevitable to achieve the joint balance across all covariates. To mitigate this iterative procedure, we utilize the entropy balancing weighting technique which focuses on balancing the covariates of subjects between the experimental and control groups directly and augments the current trial with the external control data via a power prior. The finite-sample properties of the proposed method are assessed via simulations in the context of asymmetrically randomized controlled trials where only a small portion of patients are randomized to the control group. Other methods such as covariate-balancing propensity score (CBPS) and propensity score matching (PSM) and weighting (PSW) are also compared to provide context on the operating characteristics of the proposed method.

Repigmentation by combined narrow­band ultraviolet B/adipose­derived stem cell transplantation in the mouse model: Role of Nrf2/HO­1­mediated Ca2+ homeostasis.

Vitiligo is a depigmentation disease commonly seen in clinical practice, mainly involving loss of functional epidermal pigment cells and hair follicle melanocytes. Narrow­band ultraviolet B (NB­UVB) has emerged as the first choice of treatment for vitiligo, but long­term exposure may have serious consequences. Recently, it was reported that adipose­derived stem cells (ADSCs) improve melanocyte growth and the efficacy of melanocyte transplantation. The present study aimed to examine the efficacy of NB­UVB/ADSC­transplantation combined therapy on a mouse vitiligo model and explore the underlying mechanisms by focusing on endoplasmic reticulum stress and cellular calcium (Ca2+) homeostasis. Vitiligo mice models were established by applying 40% monobenzone (MBZ) cream twice daily and treated with NB­UVB/ADSC combination therapy. Some treated mice were also given ML385, a nuclear factor erythroid 2 like 2 (Nr2) inhibitor. Histopathological changes were evaluated using a depigmentation evaluation score and observed with hematoxylin and eosin staining on skin tissues. ELISA was used to measure diagnostic markers in plasma. Flow cytometric assay was performed to quantify CD3+, CD4+ and CD8+ levels. Expression levels of associated proteins were detected with western blot and immunofluorescence. Treatment of mice with MBZ­induced depigmentation patches on the skin was accompanied with loss of redox balance and disruption of cellular Ca2+ homeostasis. Oxidative stress and Ca2+ unbalancing were improved after the mice were treated by NB­UVB/ADSCs transplantation combination therapy. ML385, strongly negated the protective effect of NB­UVB/ADSC transplantation combination therapy, indicating the critical role of Nr2 signaling. The findings improved the understanding of the pathogenesis of vitiligo and will guide future development of therapeutic strategies against it.

Discovery of 1-(5-(1H-benzo[d]imidazole-2-yl)-2,4-dimethyl-1H-pyrrol-3-yl)ethan-1-one derivatives as novel and potent bromodomain and extra-terminal (BET) inhibitors with anticancer efficacy.

As epigenetic readers, bromodomain and extra-terminal domain (BET) family proteins bind to acetylated-lysine residues in histones and recruit protein complexes to promote transcription initiation and elongation. Inhibition of BET bromodomains by small molecule inhibitors has emerged as a promising therapeutic strategy for cancer. Herein, we describe our efforts toward the discovery of a novel series of 1-(5-(1H-benzo[d]imidazole-2-yl)-2,4-dimethyl-1H-pyrrol-3-yl)ethan-1-one derivatives as BET inhibitors. Intensive structural modifications led to the identification of compound 35f as the most active inhibitor of BET BRD4 with selectivity against BET family proteins. Further biological studies revealed that compound 35f can arrest the cell cycle in G0/G1 phase and induce apoptosis via decreasing the expression of c-Myc and other proteins related to cell cycle and apoptosis. More importantly, compound 35f showed favorable pharmacokinetic properties and antitumor efficacy in MV4-11 mouse xenograft model with acceptable tolerability. These results indicated that BET inhibitors could be potentially used to treat hematologic malignancies and some solid tumors.

Age-dependent effects of transverse maxillary deficiency treated by microimplant-assisted rapid palatal expansion: A prospective cone-beam computed tomography study.

INTRODUCTION: This study investigated the results of maxillary discrepancy treated by microimplant-assisted rapid palatal expansion (MARPE) at different ages. METHODS: Sixty patients (aged 11.0-34.1 years; 23 male and 37 female) were treated by MARPE. Cone-beam computed tomography scans and dental casts were taken before and after expansion. The data were compared among 4 age groups: early adolescents (aged 11-14 years), late adolescents (aged 15-19 years), young adults (aged 20-24 years), and old adults (aged 25-34.1 years). RESULTS: The success rates of midpalatal suture separation were 100%, 100%, 88.2%, and 85.7% for early adolescents, late adolescents, young adults, and old adults, respectively. Palatal sutures at the level of the first molar were expanded by 4.02 mm, 3.48 mm, 2.63 mm, and 2.10 mm, corresponding to 66.7%, 58.1%, 42.0%, and 37.9% of the total dental expansion. Significant differences were found in the amounts of palatal suture expansion and the ratio of skeletal dental expansion between patients aged <20 years and patients aged ≥20 years (P <0.05). Skeletal expansion constituted 69.4%, 51.3%, 39.0%, and 29.8% of the total screw expansion. Except for the comparison between young adults and old adults, there were significant differences between groups for the ratio of skeletal screw expansion (P <0.05). CONCLUSIONS: The midpalatal suture can be expanded by MARPE more easily in patients <20 years of age than in patients ≥20 years of age. The ratio of skeletal screw expansion decreases as age increases.

Synergistic effect of docosahexaenoic acid or conjugated linoleic acid with caffeic acid on ameliorating oxidative stress of HepG2 cells.

Exploring the synergistic effect of docosahexaenoic acid (DHA) or conjugated linoleic acid (CLA) with caffeic acid (CA) on ameliorating oxidative stress, thereby introducing CA to DHA or CLA will contribute significantly to enhance the bioactivity. We observed that DHA or CLA with CA promoted the recovery of intact individual morphology and the decline of cavities inside the nucleus and apoptosis under the observation of confocal laser scanning microscopy and fluorescent inverted microscope. The activity of intracellular antioxidant enzymes catalase (CAT) and glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH) leakage, pyruvate and malondialdehyde and reactive oxygen species (ROS), cellular morphology, and cell cycle were analyzed. Our results showed that DHA or CLA with CA enhanced the activity of CAT and GSH-Px, decreased LDH leakage and the number of apoptotic, significantly inhibited (ROS-induced cellular injury. Cell arrest in G1 and G2 phase during cell mitosis was reduced by the measurement of flow cytometry. DHA or CLA combined with CA could markedly strengthen the free radical scavenging and endogenous antioxidant defense capacity on HepG2 cells. This study provides a new direction in the application of synergies to antioxidant compounds. PRACTICAL APPLICATION: Caffeic acid (CA) can synergize with docosahexaenoic acid (DHA) or conjugated linoleic acid (CLA) to enhance antioxidant capacity. This study highlighted an effect of ameliorating oxidative stress injury DHA or CLA with CA on HepG2 cells. The data indicated that DHA or CLA with CA might be used to relieve oxidative stress damage.

Simvastatin intervention mitigates hypercholesterolemia-induced alveolar bone resorption in rats.

Simvastatin promotes bone formation and increases bone mineral density in patients with hyperlipidemia and ameliorates hypercholesterolemia-induced microstructure changes in the jaw bone of animals. However, whether and how treatment with simvastatin can modulate the hypercholesterolemia-induced alveolar bone resorption is unclear. The present study aimed to examine the therapeutic efficacy and potential mechanisms of simvastatin application in hypercholesterolemia-induced alveolar bone resorption. The association between hyperlipidemia and alveolar bone resorption in 100 patients with periodontitis was examined. Additionally, male Sprague-Dawley rats were fed a standard rodent chow (NC) for 32 weeks or a high cholesterol diet (HCD) for 24 weeks. The HCD-fed rats were randomized, continually fed with HCD and treated with vehicle saline (HC) or simvastatin by gavage (5 mg/kg; SIM, n=10/group) for 8 weeks. The morphological changes to alveolar bone resorption in rats were analyzed by linear measurements. The relative levels of osteoprotegerin (OPG), receptor activator of nuclear factor-κB ligand RANKL, nuclear factor-κB (NF-κB), microtubule-associated protein 1 light chain 3 (LC3) and p62 in the alveolar bone tissues were determined by reverse transcription-quantitative PCR and/or immunohistochemistry. Sulcus bleeding index (SBI), clinical attachment loss (CAL), probing depth (PD) and the distance of cemantoenamel junction-alveolar bone crest (CEJ-ABC) in patients with hyperlipidemia were significantly greater than that in the controls (P<0.001). The levels of hyperlipidemia were positively correlated with the values of SBI, CAL, PD and CEJ-ABC in this population. Compared with the NC rats, higher levels of alveolar bone resorption, NF-κB expression, higher ratios of RANKL/OPG mRNA transcripts and LC3 to p62 expression were detected in the alveolar bone tissues of HC group. Simvastatin intervention significantly mitigated hypercholesterolemia-induced alveolar bone loss and RANKL mRNA transcription, but increased the ratios of LC3/p62 protein expression in the alveolar bone tissues of rats. Hyperlipidemia is associated with alveolar bone resorption and simvastatin treatment alleviated the hypercholesterolemia-related alveolar bone loss by down-regulating the NF-κB expression.

SPA: Single patient acceleration in oncology dose-escalation trials.

Efficient identification of the optimal dose and dosing scheme is one of the most critical and challenging tasks in early-phase oncology trials. The results are far-reaching because advancing a sub-optimal dose to late-stage development may not only jeopardize patients' safety or fail to deliver desired efficacy, but also be costly to sponsors as refined doses must be evaluated further before seeking regulatory approval. A good dose-escalation design is anticipated to yield high accuracy of selecting the correct dose while using fewer patients and keeping the trial duration short. Recently, treating a single patient at each lower dose level until certain events are triggered to switch to larger cohorts has gained much popularity. We name this approach "Single Patient Acceleration" (SPA), which is essentially a variant of the Accelerated Titration Design (ATD) by Simon et al. [25]. Although literature on novel dose-escalation methods is abundant in the past decade, there is a surprisingly lack of research on evaluating the ATD/SPA framework. In this article, we conduct comprehensive simulations to evaluate the performance of dose-escalation designs with or without SPA, and show that SPA improves design efficiency with similar or better accuracy to those without the "single patient" component under certain circumstances (e.g., slow initial enrollment, or the true maximum tolerated dose is at higher candidate dose levels). Potential safety concerns as a cost of efficiency improvement are also investigated in a quantitative manner to illustrate a comprehensive benefit-risk profile of SPA. Practical considerations and recommendations in using SPA are also discussed.

Comparison of skeletal maxillary transverse deficiency treated by microimplant-assisted rapid palatal expansion and tooth-borne expansion during the post-pubertal growth spurt stage.

OBJECTIVE: To investigate the efficacy of microimplant-assisted rapid palatal expansion (MARPE) to treat skeletal maxillary discrepancies during the post-pubertal growth spurt stage. MATERIALS AND METHODS: Sixty patients with skeletal maxillary transverse deficiency during the post-pubertal growth spurt stage were randomly divided into MARPE and Hyrax groups. Thirty patients (mean age: 15.1 ± 1.6 years) were treated using the four-point MARPE appliance; 30 patients (mean age, 14.8 ± 1.5 years) were treated using the Hyrax expander. Cone beam computed tomography scans and dental casts were obtained before and after expansion. The data were analyzed using paired t-tests and independent t-tests. RESULTS: The success rates of midpalatal suture separation were 100% and 86.7% for MARPE and Hyrax groups, respectively. Palatal expansion and skeletal to dental ratio at the first molar level were greater in the MARPE group (3.82 mm and 61.4%, respectively) than in the Hyrax group (2.20 mm and 32.3%, respectively) (P < .01). Reductions in buccal alveolar bone height and buccal tipping of the first molars were less in the MARPE group than in the Hyrax group (P < .01). CONCLUSIONS: MARPE enabled more predictable and greater skeletal expansion, as well as less buccal tipping and alveolar height loss on anchorage teeth. Thus, MARPE is a better alternative for patients with skeletal maxillary deficiency during the post-pubertal growth spurt stage.

Changing Expression Profiles and Inclination to Competing Endogenous RNA Networks on MAPK Signaling Pathways of Human Adipose-Derived Stem Cells in a Direct Current Electric Field.

Adipose-derived stem cells (ADSCs) are an abundant cell source and provide an easy way to harvest mesenchymal stem cells, which are the focus of considerable attention in regenerative medicine. Electric fields (EF) play roles in many biological events and have been reported to promote cell proliferation, migration, and differentiation. In this study, ADSCs were treated with a direct current electric field (DCEF) of either 0 (control group) or 300 mV/mm (EF group) for six hours. RNA screening and analysis revealed that 66, 164, 26, and 1310 circRNAs, lncRNAs, miRNAs, and mRNAs, respectively, were differentially expressed in the DCEF-treated ADSCs compared with untreated ADSCs. Differentially expressed mRNAs were enriched in the MAPK signaling pathway, TNF signaling pathway, and some other pathways. ANXA1, ERRFI1, JAG1, EPHA2, PRR9, and H2AFY2 were related to the keratinocyte differentiation process. Competing endogenous RNA (ceRNA) networks were constructed on the basis of genes in the MAPK signaling pathway. Twenty-one RNAs in the above networks were randomly chosen, and their expression was validated using qRT-PCR, which showed the same expression trends as the RNA sequencing analysis. The MAPK signaling pathway is of great importance in the ADSC processes that occur in a DCEF, including keratinocyte differentiation. Several ceRNAs may participate in the regulation of MAPK signaling. This study may give new insight into the proliferation, migration, and differentiation of ADSCs, which will be valuable for tissue engineering and regenerative medicine.

PairedFB: a full hierarchical Bayesian model for paired RNA-seq data with heterogeneous treatment effects.

MOTIVATION: Several methods have been proposed for the paired RNA-seq analysis. However, many of them do not consider the heterogeneity in treatment effect among pairs that can naturally arise in real data. In addition, it has been reported in literature that the false discovery rate (FDR) control of some popular methods has been problematic. In this paper, we present a full hierarchical Bayesian model for the paired RNA-seq count data that accounts for variation of treatment effects among pairs and controls the FDR through the posterior expected FDR. RESULTS: Our simulation studies show that most competing methods can have highly inflated FDR for small to moderate sample sizes while PairedFB is able to control FDR close to the nominal levels. Furthermore, PairedFB has overall better performance in ranking true differentially expressed genes (DEGs) on the top than others, especially when the sample size gets bigger or when the heterogeneity level of treatment effects is high. In addition, PairedFB can be applied to identify the biologically significant DEGs with controlled FDR. The real data analysis also indicates PairedFB tends to find more biologically relevant genes even when the sample size is small. PairedFB is also shown to be robust with respect to the model misspecification in terms of its relative performance compared to others. AVAILABILITY AND IMPLEMENTATION: Software to implement this method (PairedFB) can be downloaded at: https://sites.google.com/a/udel.edu/qiujing/publication. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

miR-17-5p promotes proliferation and migration of CAL-27 human tongue squamous cell carcinoma cells involved in autophagy inhibition under hypoxia.

Autophagy contributes to head and neck squamous cell carcinoma (HNSCC) development and progression. MiR-17-5p down-regulates Beclin-1 and thus plays an important role in autophagy, but little is known about the function and regulation of miR-17-5p in HNSCC autophagy. This study aimed to investigate the role of miR-17-5p on proliferation, migration, and autophagy under hypoxia in CAL-27 human tongue squamous cell carcinoma cells. CAL-27 cells were transfected with 50 nmol miR-17-5p mimics to overexpress miR-17-5p. Cell proliferation and migration were determined by CCK-8 and wound healing assays, respectively, under hypoxia. Autophagy induced by hypoxia was detected by transmission electron microscope and Beclin-1 mRNA and protein expressions. The miR-17-5p mimics successfully increased the expression of miR-17-5p in CAL-27 cells by almost 700 fold compared with the miRNA mimic negative control. After 3 days, cells transfected with the miR-17-5p mimics showed higher proliferation compared with controls (P < 0.05) under hypoxia. MiR-17-5p transfected CAL-27 cells had a higher migratory capacity compared with the control cells (P < 0.05) under hypoxia. Furthermore, transmission electron microscopy showed that miR-17-5p overexpression inhibited the formation of autophagosomes in hypoxic cells. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot showed that miR-17-5p overexpression inhibited the mRNA and protein expression of Beclin-1 in CAL-27 cells submitted to hypoxia. MiR-17-5p overexpression promoted the proliferation and migration of the CAL-27 cells, but inhibited autophagy under hypoxia.

Exact test-based approach for equivalence test with parameter margin.

The equivalence test has a wide range of applications in pharmaceutical statistics which we need to test for the similarity between two groups. In recent years, the equivalence test has been used in assessing the analytical similarity between a proposed biosimilar product and a reference product. More specifically, the mean values of the two products for a given quality attribute are compared against an equivalence margin in the form of ±f × σR, where ± f × σ R is a function of the reference variability. In practice, this margin is unknown and is estimated from the sample as ±f × SR. If we use this estimated margin with the classic t-test statistic on the equivalence test for the means, both Type I and Type II error rates may inflate. To resolve this issue, we develop an exact-based test method and compare this method with other proposed methods, such as the Wald test, the constrained Wald test, and the Generalized Pivotal Quantity (GPQ) in terms of Type I error rate and power. Application of those methods on data analysis is also provided in this paper. This work focuses on the development and discussion of the general statistical methodology and is not limited to the application of analytical similarity.

Targeting epigenetic reader and eraser: Rational design, synthesis and in vitro evaluation of dimethylisoxazoles derivatives as BRD4/HDAC dual inhibitors.

The bromodomain protein module and histone deacetylase (HDAC), which recognize and remove acetylated lysine, respectively, have emerged as important epigenetic therapeutic targets in cancer treatments. Herein we presented a novel design approach for cancer drug development by combination of bromodomain and HDAC inhibitory activity in one molecule. The designed compounds were synthesized which showed inhibitory activity against bromodomain 4 and HDAC1. The representative dual bromodomain/HDAC inhibitors, compound 11 and 12, showed potent antiproliferative activities against human leukaemia cell line K562 and MV4-11 in cellular assays. This work may lay the foundation for developing dual bromodomain/HDAC inhibitors as potential anticancer therapeutics.