Self-Report Amphiphilic Polymer-Based Drug Delivery System with ROS-Triggered Drug Release for Osteoarthritis Therapy.

The development of drug delivery systems with real-time cargo release monitoring capabilities is imperative for optimizing nanomedicine performance. Herein, we report an innovative self-reporting drug delivery platform based on a ROS-responsive random copolymer (P1) capable of visualizing cargo release kinetics via the activation of an integrated fluorophore. P1 was synthesized by copolymerization of pinacol boronate, PEG, and naphthalimide monomers to impart ROS-sensitivity, hydrophilicity, and fluorescence signaling, respectively. Detailed characterization verified that P1 self-assembles into 11 nm micelles with 10 µg mL-1 CMC and can encapsulate hydrophobic curcumin with 79% efficiency. Fluorescence assays demonstrated H2O2-triggered disassembly and curcumin release with concurrent polymer fluorescence turn-on. Both in vitro and in vivo studies validated the real-time visualization of drug release and ROS scavenging, as well as the therapeutic effect on osteoarthritis (OA). Overall, this nanotheranostic polymeric micelle system enables quantitative monitoring of drug release kinetics for enhanced treatment optimization across oxidative stress-related diseases.

Sensitive Imaging of Cellular RNA via Cascaded Proximity-Induced Fluorogenic Reactions.

Owing to its important biological functions, RNA has become a promising molecular biomarker of various diseases. With a dynamic change in its expression level and a relatively low amount within the complicated biological matrix, signal amplification detection based on DNA probes has been put forward, which is helpful for early diagnosis and prognostic prediction. However, conventional methods are confined to cell lysates or dead cells and are not only time-consuming in sample preparation but also inaccessible to the spatial-temporal information of target RNAs. To achieve live-cell imaging of specific RNAs, both the detection sensitivity and intracellular delivery issues should be addressed. Herein, a new cascaded fluorogenic system based on the combination of hybridization chain reactions (HCRs) and proximity-induced bioorthogonal chemistry is developed, in which a bioorthogonal reaction pair (a tetrazine-quenched dye and its complementary dienophile) is brought into spatial proximity upon target RNA triggering the HCR to turn on and amplify the fluorescence in one step, sensitively indicating the cellular distribution of RNA with minimal false positive results caused by unspecific degradation. Facilitated by a biodegradable carrier based on black phosphorus with high loading capacity and excellent biocompatibility, the resulting imaging platform allows wash-free tracking of target RNAs inside living cells.

Iron overload promotes intervertebral disc degeneration via inducing oxidative stress and ferroptosis in endplate chondrocytes.

Iron overload is a common phenomenon in the elderly population. Many clinical studies have indicated an association between iron overload and the incidence and pathological progression of intervertebral disc degeneration (IVDD). However, the role and underlying mechanism by which iron participates in the progression of IVDD has not yet been reported. In the present study, we aimed to elucidate the connection between iron overload and IVDD, and explore the underlying mechanisms of disease. Firstly, a clinical epidemiology study was conducted and revealed that iron overload is an independent risk factor for human IVDD. To elucidate the role of iron overload in IVDD, an iron overload mouse model was established, and we observed that iron overload promoted IVDD and cartilage endplate degeneration in a dose dependent manner. Endplate chondrocytes were further isolated and treated with FAC to mimic iron overload in vitro. Excess iron significantly promoted mineralization of endplate chondrocytes in addition to their degeneration via oxidative stress. Moreover, a high dose of excess iron promoted chondrocytes ferroptosis. An iron chelator (DFO), an antioxidant (NAC) and a ferroptosis inhibitor (Fer-1) demonstrated effective inhibition of endplate chondrocyte degeneration induced by iron overload, and our in vivo studies further demonstrated that DFO, NAC and Fer-1 could rescue high dose iron-induced IVDD and cartilage endplate calcification. In conclusion, our results indicate that iron overload is strongly associated with the onset and development of IVDD via oxidative stress and ferroptosis. Inhibiting oxidative stress or ferroptosis could therefore be promising therapeutic strategies for IVDD induced by iron overload.

Mindfulness Moderates the Association Between Perceived Discrimination and Cyber Aggression Among Emerging Adults with Early Left-Behind Experience: A Longitudinal Study.

Purpose: Although the risk effect of perceived discrimination on left-behind youth's mental health (mainly emotional problems) has been demonstrated in prior studies, there is a lack of longitudinal studies examining the effect of perceived discrimination on behavioral problems, particularly in emerging adults with early left-behind experience. In addition, little is known about individual differences in terms of this association. In the present study, we draw on the social information processing model of aggression to examine the effect of perceived discrimination and mindfulness on cyber aggression. Methods: We used two-wave longitudinal panel data involving 535 emerging adults with early left-behind experience in rural China (M age = 19.89 years; SD = 1.20; 57.2% female). To test the moderating role of mindfulness, hierarchical multiple regression and simple slope test analyses were performed. Results: The results of linear regression analysis demonstrated that higher levels of perceived discrimination at T1 were significantly associated with higher cyber aggression at T2; the strength of this association was buffered by mindfulness over time. Conclusion: These findings suggest that the facilitation of mindfulness training may be an effective strategy for reducing the risk of perceived discrimination leading to cyber aggression in emerging adults with early left-behind experience.

An overview of kinase downregulators and recent advances in discovery approaches.

Since the clinical approval of imatinib, the discovery of protein kinase downregulators entered a prosperous age. However, challenges still exist in the discovery of kinase downregulator drugs, such as the high failure rate during development, side effects, and drug-resistance problems. With the progress made through multidisciplinary efforts, an increasing number of new approaches have been applied to solve the above problems during the discovery process of kinase downregulators. In terms of in vitro and in vivo drug evaluation, progress was also made in cellular and animal model platforms for better and more clinically relevant drug assessment. Here, we review the advances in drug design strategies, drug property evaluation technologies, and efficacy evaluation models and technologies. Finally, we discuss the challenges and perspectives in the development of kinase downregulator drugs.

Blocking exposed PD-L1 elicited by nanosecond pulsed electric field reverses dysfunction of CD8+ T cells in liver cancer.

As a promising method for local tumor treatment, nanosecond pulsed electric field (nsPEF) ablation elicits a potent anti-tumor immune response. However, the mechanism of the nsPEF-mediated anti-tumor immune response and its effects on the tumor microenvironment remains unclear. Here, we demonstrated that nsPEF treatment increased the level of membrane PD-L1 in liver cancer cells. Furthermore, nsPEF induced the release of PD-L1-associated extra-cellular vesicles, leading to the dysfunction of CD8+ T cells, which could potentially be reversed by PD-L1 blockade. Biological and functional assays also demonstrated that nsPEF treatment resulted in the increased PD-L1 level and dysfunction of infiltrated CD8+ T cells in tumor tissues in vivo, indicating the long term antitumor efficacy of nsPEF treatment. A combination of nsPEF treatment and PD-L1 blockade effectively inhibited tumor growth and improved the survival of the tumor-bearing mouse. In conclusion, nsPEF treatment induced the translocation and release of PD-L1 and contributed to the dysfunction of infiltrated CD8+ T cells, resulting in tumor progression at later stages. The combination of nsPEF treatment and PD-L1 blockade is a promising therapeutic strategy for liver cancer.

Network-based analysis with primary cells reveals drug response landscape of acute myeloid leukemia.

Acute myeloid leukemia (AML) is one of the most common, complex, and heterogeneous hematological malignancies in adults. Despite progresses in understanding the pathology of AML, the 5-year survival rates still remain low compared with CML, CLL, etc. The relationship between genomic features and drug responses is critical for precision medication. Herein, we depicted a picture for response of 145 drugs against 33 primary cell samples derived from AML patients with full spectrum of genomic features assessed by whole exon sequencing and RNA sequencing. In general, most of the samples were much more sensitive to the combinatorial chemotherapy regimens than the single chemotherapy drugs. Overall, these samples were moderately sensitive to the Traditional Chinese Medicine (TCM) and the targeted drugs. In the weighted gene coexpression network analysis (WGCNA), the TCM and targeted therapies displayed similar genetic signatures in the gene module correlation. Meanwhile, the expression of miRNAs, lncRNAs, and mRNAs did not display apparent gene module correlations among those different types of therapies. In addition, the combinatorial chemotherapy bear more module correlations than the single drugs. Interestingly, we found that the gene mutations and drug response were not enriched in any WGCNA module analysis. Most of the sensitive drug response biomarkers were enriched in the ribosome, endocytosis, cell cycle, and p53 associated signaling pathways. This study showed that gene expression modules might show better correlation than gene mutations for drug efficacy predictions.

Discovery of N-(4-(6-Acetamidopyrimidin-4-yloxy)phenyl)-2-(2-(trifluoromethyl)phenyl)acetamide (CHMFL-FLT3-335) as a Potent FMS-like Tyrosine Kinase 3 Internal Tandem Duplication (FLT3-ITD) Mutant Selective Inhibitor for Acute Myeloid Leukemia.

Most of the current FMS-like tyrosine kinase 3 (FLT3) inhibitors lack selectivity between FLT3 kinase and cKIT kinase as well as the FLT3 wt and internal tandem duplication (ITD) mutants. We report a new compound 27, which displays GI50 values of 30-80 nM against different ITD mutants and achieves selectivity over both FLT3 wt (8-fold) and cKIT kinase in the transformed BaF3 cells (>300-fold). 27 potently inhibits the proliferation of the FLT3-ITD-positive acute myeloid leukemia cancer lines through suppression of the phosphorylation of FLT3 kinase and downstream signaling pathways, induction of apoptosis, and arresting the cell cycle into the G0/G1 phase. 27 also displays potent antiproliferative effect against FLT3-ITD-positive patient primary cells, whereas it does not apparently affect FLT3 wt primary cells. In addition, it also exhibits a good therapeutic window to PBMC compared to PKC412. In the in vivo studies, 27 demonstrates favorable PK profiles and suppresses the tumor growth in the MV4-11 cell inoculated mouse xenograft model.

Remediation of lead-contaminated water by geological fluorapatite and fungus Penicillium oxalicum.

Phosphate-solubilizing fungi (PSF) can secrete large amounts of organic acids. In this study, the application of the fungus Penicillium oxalicum and geological fluorapatite (FAp) to lead immobilization was investigated. The formation and morphology of the lead-related minerals were analyzed by ATR-IR, XRD, Raman, and SEM. The quantity of organic acids secreted by P. oxalicum reached the maximum on the fourth day, which elevated soluble P concentrations from 0.4 to 108 mg/L in water. The secreted oxalic acid dominates the acidity in solution. P. oxalicum can survive in the solution with Pb concentration of ~ 1700 mg/L. In addition, it was shown that ~ 98% lead cations were removed while the fungus was cultured with Pb (~ 1700 mg/L) and FAp. The mechanism is that the released P from FAp (enhanced by organic acids) can react with Pb2+ to form the stable pyromorphite mineral [Pb5(PO4)3F]. The precipitation of lead oxalate also contributes to Pb immobilization. However, lead oxalate is more soluble due to its relatively high solubility. P. oxalicum has a higher rate of organic acid secretion compared with other typical PSF, e.g., Aspergillus niger. This study sheds light on bright future of applying P. oxalicum in Pb remediation.

Moderate intensity static magnetic fields affect mitotic spindles and increase the antitumor efficacy of 5-FU and Taxol.

Microtubules are the fundamental components in mitotic spindle, which plays essential roles in cell division. It was well known that purified microtubules could be affected by static magnetic fields (SMFs) in vitro because of the diamagnetic anisotropy of tubulin. However, whether these effects lead to cell division defects was unknown. Here we find that 1T SMFs induce abnormal mitotic spindles and increase mitotic index. Synchronization experiments show that SMFs delay cell exit from mitosis and cause mitotic arrest. These mimic the cellular effects of a microtubule-targeting drug Paclitaxel (Taxol), which is frequently used in combination with 5-Fluorouracil (5-FU) and Cisplatin in cancer treatment. Using four different human cancer cell lines, HeLa, HCT116, CNE-2Z and MCF7, we find that SMFs increase the antitumor efficacy of 5-FU or 5-FU/Taxol, but not Cisplatin, which indicates that the SMF-induced combinational effects with chemodrugs are drug-specific. Our study not only reveals the effect of SMFs on microtubules to cause abnormal mitotic spindles and delay cells exit from mitosis, but also implies the potential applications of SMFs in combination with chemotherapy drugs 5-FU or 5-FU/Taxol, but not with Cisplatin in cancer treatment.

The biological characteristics of a novel camptothecin-artesunate conjugate.

A novel conjugate of camptothecin and artesunate (C-Q) was prepared and its cytotoxicity was evaluated using the MTT assay. In addition, the antitumour activity and toxicity of C-Q were investigated in mice, and interaction between transferrin (TF) and C-Q was investigated to evaluate its interaction with biological macromolecules. In the MTT assay, C-Q showed better inhibitory activity against MCF7 breast cancer cells and SMMC-7721 liver cancer cells than camptothecin or artesunate. In vivo, C-Q showed lower toxicity and better antitumour activity compared with camptothecin. Fluorescence spectroscopy showed static quenching of TF in the presence of C-Q, and thermodynamic parameters (ΔH>0 and ΔG<0) indicated that the reaction was spontaneous and endothermic. The main binding force between C-Q and TF was hydrophobic, as indicated by thermodynamic parameters (ΔH>0 and ΔS>0). Thus, synchronous fluorescence spectra showed that C-Q had no influence on the conformation of TF. Our results indicated that C-Q represents a novel potential anticancer therapeutic vector with advantages over current methods of CPT and ART administration. This novel drug delivery system allows the use of these drugs in a manner associated with few side effects for normal tissue, and which facilitates synergistic effects of anti-tumour drugs.

Tai Chi exercise versus rehabilitation for the elderly with cerebral vascular disorder: a single-blinded randomized controlled trial.

BACKGROUND: Cerebral vascular disorder (CVD) might result in a quantifiable decrease in quality of life, which is determined not only by the neurological deficits but also by impairment of cognitive functions. There are few studies that report on the cognitive effect of Tai Chi exercise (Tai Chi) on the elderly with CVD. The purpose of the present study was to examine the cognitive effect of Tai Chi on the elderly with CVD using P300 measurement, in addition to the General Health Questionnaire (GHQ) and Pittsburgh Sleep Quality Index (PSQI). METHODS: A total of 34 patients with CVD were recruited from outpatient Akistu-Kounoike Hospital and randomly assigned to receive Tai Chi (n= 17) or rehabilitation (n= 17) in group sessions once a week for 12 weeks. To examine the time courses of each score (P300 amplitude, P300 latency, GHQ score and PSQI score), repeated-measures analysis of variance was carried out with groups and time as factors. RESULTS: For the time courses of P300 amplitudes and latencies, there were no significant effects of interaction between group and time. However, significant time-by-group interactions were found for Sleep Quality (P= 0.006), GHQ total score (P= 0.005), anxiety/insomnia score (P= 0.034), and severe depression score (P= 0.020). CONCLUSIONS: Tai Chi might therefore be considered a useful non-pharmacological approach, along with rehabilitation, for the maintenance of cognitive function in the elderly with CVD and might be a more useful non-pharmacological approach for the improvement of sleep quality and depressive symptoms in the elderly with CVD than rehabilitation.

Visualized regional hypodermic migration channels of interstitial fluid in human beings: are these ancient meridians?

BACKGROUND: Since the 1980s, many studies have indicated there are hypodermic migration channels of radiotracer along meridians independent of lymphatic and blood vessels in vivo. However, the radioactive trajectories were not clear enough to identify the anatomic structures in the living body. OBJECTIVES: To establish an alternative method to image the specific fluid pathways, we hypothesized that a minimal amount of low molecular weight tracer could enter the specific channels and by magnetic resonance imaging technique, we could image the regional channels originating from the acupoints on 6 yin meridians of forearm and lower leg. DESIGN AND RESULTS: Seven (7) healthy volunteers were injected hypodermically with a minimal amount of tracer into 6 acupoints on 6 yin meridians of the hand and foot. We found a total of 6 regional specific channels in one forearm and one lower leg of each subject, respectively, but no specific channels were visualized following injection of the tracer into the sites of nonacupoints. Magnetic resonance angiography confirmed the specific channels were not the superficial veins in the subcutis. Subsequent acupuncture needling studies revealed that migration of tracer along each of the specific channels could not be interrupted by acupuncture needling and remained intact, which was different from the nature of either lymphatics or blood vessels. CONCLUSIONS: We imaged 6 regional migration channels originating from 6 acupoints on 6 yin meridians of hand and foot by injecting a minimal amount of tracer into acupoints directly in humans. The responses of the subcutaneous specific channels to acupuncture needling are different from those of lymphatic or blood vessels and coincide partially with the characteristics of acupuncture therapeutics, suggesting the existence of meridian-like channels in humans. These findings may contribute to further research on the specific interstitial fluid drainage.

Functional magnetic resonance imaging and immunohistochemical study of hypothalamic function following oral glucose ingestion in rats.

BACKGROUND: The hypothalamus plays a central role in the regulation of metabolism by sensing metabolic demands and releasing regulatory neurotransmitters. This study investigated the response of the hypothalamus to glucose ingestion in rats by blood oxygen level-dependent functional magnetic resonance imaging (BOLD-fMRI) and immunohistochemical techniques to determine the role of the hypothalamus in glyco-regulation during disturbances in carbohydrate metabolism. METHODS: The signal intensity of the hypothalamus was monitored by fMRI for 60 minutes after oral glucose intake in 48 healthy rats (age 14 months), which included 24 normal weight rats (weighing (365 +/- 76.5) g) and 24 overweight rats (weighing (714 +/- 83.5) g). Then, 12 rats (6 normal, 6 overweight) underwent a repeat fMRI scan after consuming an equivalent amount of water without glucose on a separate day. The procedure for fMRI with water intake was the same as for glucose ingestion. fMRI data was processed using time cluster analysis and intensity averaging method. After fMRI, the expression of neuropeptide Y (NPY) and 5-hydroxytryptamine (5-HT) in the hypothalamus of all rats was determined by immunohistochemistry. Positive cells for NPY or 5-HT were counted. RESULTS: There was a transient, but significant, decrease in fMRI signal intensity in all rats (mean (3.12 +/- 0.78)%) in the hypothalamus within 19.5 - 25.5 minutes of oral glucose ingestion. In overweight rats, the decrease in signal intensity in response to the glucose ingestion was more markedly attenuated than that observed in normal weight rats ((2.2 +/- 1.5)% vs (4.2 +/- 0.7)% inhibition, t = 2.12, P < 0.05). There was no significant response in the hypothalamus after oral water ingestion. The percentage of NPY positive cells in obese rats were slightly lower than those in control group (21% vs 23%, t = 0.71, P > 0.05); but there was no significant difference between the two groups; the percentage of 5-HT positive cells in obese rats were significantly lower than those in the control group (22% vs 31%, t = 3.25, P < 0.01). CONCLUSIONS: There is a transient, but significant, decrease in BOLD signal intensity in the hypothalamus following glucose ingestion, which is similar to that observed in humans. The response of the hypothalamus to glucose ingestion was different in overweight and normal weight rats. The percentage of NPY positive cells in obese rats were lower than those in the control group, although this difference was not statistically significant. The percentage of 5-HT positive cells in obese rats was significantly lower than those in the control group.