Genetically Engineered Biomimetic Nanoparticles for Targeted Delivery of mRNA to Treat Rheumatoid Arthritis.

In addition to inhibiting persistent inflammation, phosphatase and tensin homolog deleted from chromosome 10 (PTEN) is known as an important therapeutic target for alleviating rheumatoid arthritis (RA) symptoms. Modulation of PTEN gene expression in synovial tissue using messenger RNA (mRNA) is a promising approach to combat RA. However, mRNA therapeutics are often hampered by unsatisfactory stability and inefficient localization in synovial tissue. In this study, a genetically engineered biomimetic membrane-coated mRNA (MR@P-mPTEN) carrier that effectively delivers mRNA-PTEN (mPTEN) directly to the RA joint is presented. By overexpressing tumor necrosis factor (TNF-α) receptors on macrophage biomimetic membranes via plasmid transfection, decoys that reduce inflammatory pathway activation are prepared for TNF-α. The resulting construct, MR@P-mPTEN, shows good stability and RA targeting based on in vivo fluorescence imaging. It is also found that MR@P-mPTEN competitively binds TNF-α and activates the PTEN pathway in vitro and in vivo, thereby inhibiting synovitis and joint damage. Clinical micro-computed tomography and histological analyses confirm the treatment effects. These results suggest that the genetically engineered biomimetic therapeutic platform MR@P-mPTEN both inhibits pro-inflammatory cytokines and upregulates PTEN protein expression to alleviate RA damage, providing a new a new combination strategy for RA treatment.

HIF-1α dependent RhoA as a novel therapeutic target to regulate rheumatoid arthritis fibroblast-like synoviocytes migration in vitro and in vivo.

Objective: The purpose of this work is to investigate how the Rho family of GTPases A (RhoA) mediates the pathogenesis of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS). Methods: The expression of RhoA in the synovial tissues of RA and Healthy people (Control) was detected using immunohistochemistry methods. The expression of RhoA and hypoxia-inducible factor-1α (HIF-1α) is inhibited by small interfering RNAs (siRNAs). The inhibition effect on RA-FLS migration was further investigated. The protein expression level of HIF-1α, RhoA, focal adhesion kinase (FAK), and myosin light chain (MLC) was also analysed using western blotting (WB). DBA1 mice were immunised with the mixture of bovine type II collagen and Freund's adjuvant to establish collagen induced arthritis (CIA) mouse model. Lip-siRhoA is administered through joint injection every two days. Micro-computed tomography (micro-CT) was used to detect mouse ankle joint destruction and evaluate the bone loss of the periarticular side. Destruction of the ankle articular cartilage was tested by histology. Expressions of P-RhoA, P-FAK and P-MLC in the ankle joint was detected by immunohistochemistry assay. Results: The expression level of RhoA in the synovial tissues of RA patients was significantly higher than that in control group. Hypoxia was able to up-regulate the expression of RhoA. Whereas, HIF-1α siRNA (siHIF-1α) could down-regulate the expression of RhoA. Additionally, both of siHIF-1α and RhoA siRNA (siRhoA) delivered by liposome (Lip-siHIF-1α and Lip-siRhoA) were found to suppress FAK and MLC phosphorylation in vitro. In CIA mouse model, Lip-siRhoA was demonstrated to ameliorate the destruction of ankle joint and reduce the severity of ankle joint cartilage damage by micro-CT and histological staining, respectively. Therefore, inhibition of FLS cell migration can protect articular bone from destruction. Furthermore, the expression of P-RhoA, P-FAK and P-MLC was evaluated and found to be down-regulated by Lip-siRhoA in vivo. Conclusion: The results demonstrated that under hypoxic environment, HIF-1α dependent RhoA pathway played an important role on cytoskeleton remodelling and RA-FLS migration. Through down-regulating RhoA expression, it could effectively treat RA in vitro and in vivo. The translational potential of this article: Our study provides new evidence for the potential clinical application of RhoA as a candidate for the treatment of RA.

Photoacoustic image-guided biomimetic nanoparticles targeting rheumatoid arthritis.

The high level of reactive oxygen species (ROS) in the rheumatoid arthritis (RA) microenvironment (RAM) and its persistent inflammatory nature can promote damage to joints, bones, and the synovium. Targeting strategies that integrate effective RAM regulation with imaging-based monitoring could lead to improvements in the diagnosis and treatment of RA. Here, we report the combined use of small interfering RNAs (siRNAsT/I) and Prussian blue nanoparticles (PBNPs) to silence the expression of proinflammatory cytokines TNF-α/IL-6 and scavenge the ROS associated with RAM. To enhance the in vitro and in vivo biological stability, biocompatibility, and targeting capability of the siRNAsT/I and PBNPs, macrophage membrane vesicles were used to prepare biomimetic nanoparticles, M@P-siRNAsT/I. The resulting constructs were found to suppress tumor necrosis factor-α/interleukin-6 expression and overcome the hypoxic nature of RAM, thus alleviating RA-induced joint damage in a mouse model. The M@P-siRNAsT/I of this study could be monitored via near-infrared photoacoustic (PA) imaging. Moreover, multispectral PA imaging without the need for labeling permitted the real-time evaluation of M@P-siRNAsT/I as a putative RA treatment. Clinical microcomputed tomography and histological analysis confirmed the effectiveness of the treatment. We thus suggest that macrophage-biomimetic M@P-siRNAsT/I and their analogs assisted by PA imaging could provide a new strategy for RA diagnosis, treatment, and monitoring.

Synergistic Immediate Cortical Activation on Mirror Visual Feedback Combined With a Soft Robotic Bilateral Hand Rehabilitation System: A Functional Near Infrared Spectroscopy Study.

BACKGROUND: Mirror visual feedback (MVF) has been widely used in neurological rehabilitation. Due to the potential gain effect of the MVF combination therapy, the related mechanisms still need be further analyzed. METHODS: Our self-controlled study recruited 20 healthy subjects (age 22.150 ± 2.661 years) were asked to perform four different visual feedback tasks with simultaneous functional near infrared spectroscopy (fNIRS) monitoring. The right hand of the subjects was set as the active hand (performing active movement), and the left hand was set as the observation hand (static or performing passive movement under soft robotic bilateral hand rehabilitation system). The four VF tasks were designed as RVF Task (real visual feedback task), MVF task (mirror visual feedback task), BRM task (bilateral robotic movement task), and MVF + BRM task (Mirror visual feedback combined with bilateral robotic movement task). RESULTS: The beta value of the right pre-motor cortex (PMC) of MVF task was significantly higher than the RVF task (RVF task: -0.015 ± 0.029, MVF task: 0.011 ± 0.033, P = 0.033). The beta value right primary sensorimotor cortex (SM1) in MVF + BRM task was significantly higher than MVF task (MVF task: 0.006 ± 0.040, MVF + BRM task: 0.037 ± 0.036, P = 0.016). CONCLUSION: Our study used the synchronous fNIRS to compare the immediate hemodynamics cortical activation of four visual feedback tasks in healthy subjects. The results showed the synergistic gain effect on cortical activation from MVF combined with a soft robotic bilateral hand rehabilitation system for the first time, which could be used to guide the clinical application and the future studies.

Exocyst controls exosome biogenesis via Rab11a.

Tumor cells actively release large quantities of exosomes, which pivotally participate in the regulation of cancer biology, including head and neck cancer (HNC). Exosome biogenesis and release are complex and elaborate processes that are considered to be similar to the process of exocyst-mediated vesicle delivery. By analyzing the expression of exocyst subunits and their role in patients with HNC, we aimed to identify exocyst and its functions in exosome biogenesis and investigate the molecular mechanisms underlying the regulation of exosome transport in HNC cells. We observed that exocysts were highly expressed in HNC cells and could promote exosome secretion in these cells. In addition, downregulation of exocyst expression inhibited HN4 cell proliferation by reducing exosome secretion. Interestingly, immunofluorescence and electron microscopy revealed the accumulation of multivesicular bodies (MVBs) after the knockdown of exocyst. Autophagy, the major pathway of exosome degradation, is not activated by this intracellular accumulation of MVBs, but these MVBs are consumed when autophagy is activated under the condition of cell starvation. Rab11a, a small GTPase that is involved in MVB fusion, also interacted with the exocyst. These findings suggest that the exocyst can regulate exosome biogenesis and participate in the malignant behavior of tumor cells.

Feature optimization based on improved novel global harmony search algorithm for motor imagery electroencephalogram classification.

Background: Effectively decoding electroencephalogram (EEG) pattern for specific mental tasks is a crucial topic in the development of brain-computer interface (BCI). Extracting common spatial pattern (CSP) features from motor imagery EEG signals is often highly dependent on the selection of frequency band and time interval. Therefore, optimizing frequency band and time interval would contribute to effective feature extraction and accurate EEG decoding. Objective: This study proposes an approach based on an improved novel global harmony search (INGHS) to optimize frequency-time parameters for effective CSP feature extraction. Methods: The INGHS algorithm is applied to find the optimal frequency band and temporal interval. The linear discriminant analysis and support vector machine are used for EEG pattern decoding. Extensive experimental studies are conducted on three EEG datasets to assess the effectiveness of our proposed method. Results: The average test accuracy obtained by the time-frequency parameters selected by the proposed INGHS method is slightly better than artificial bee colony (ABC) and particle swarm optimization (PSO) algorithms. Furthermore, the INGHS algorithm is superior to PSO and ABC in running time. Conclusion: These superior experimental results demonstrate that the optimal frequency band and time interval selected by the INGHS algorithm could significantly improve the decoding accuracy compared with the traditional CSP method. This method has a potential to improve the performance of MI-based BCI systems.

Analysis of risk factors of hemorrhagic transformation after acute ischemic stroke: cerebral microbleeds do not correlate with hemorrhagic transformation.

To study the potential risk factors including cerebral microbleeds (CMB) of hemorrhagic transformation (HT) after acute ischemic stroke. We included 348 consecutive patients with acute infarction who were hospitalized in two centers from June 2009 to December 2010. Acute ischemic infarctions were subdivided into atherosclerotic, cardioemblic, lacunar, and undetermined infarction groups. The related risk factors were recruited for analysis. All patients underwent gradient-echo T2-weighted imaging (GRE) to detect CMB and HT. Logistic regression analysis was used to analyze relationships, with HT as response variable and potential risk factors as explanatory variables. Multivariate logistic regression analysis demonstrated that predictor factors of HT were cardioembolic infarction (OR 24.956, 95 % CI 2.734-227.801, P = 0.004), infarction of undetermined causes (OR 19.381, 95 % CI 1.834-205.104, P = 0.014), and scores of NIHSS (OR 1.187, 95 % CI 1.109-1.292, P < 0.001), diabetes mellitus (OR 4.973, 95 % CI 2.004-12.338, P = 0.001). Whereas, the level of low-density lipoprotein was the protective factor (OR 0.654, 95 % CI 0.430-0.996, P = 0.048).The prevalence of CMB was 45.98 % (160/348) with no statistically difference among different subtypes. Thirty-five out of 348 (10.06 %) patients with ischemic stroke developed HT with a statistical difference among different subtypes of ischemia (χ (2) = 42.140, P < 0.001). The distributions of HI and PH among subgroups were variable with significant differences (χ (2) = 17.536, P = 0.001; χ (2) = 12.028, P = 0.007). PH frequency of cardioembolism was the highest (4/28, 14.29 %), and symptomatic ICH was also highest (7.14 %). The CMBs do not significantly correlate with HT. Knowledge of the risk factors associated with HT after ACI, especially HT following thrombolyitc therapy may provide insight into the mechanisms underlying the development of HT, helps to develop treatment strategy that reduces the risk of PH and implicates for the design of future acute ischemic stroke trials.