Force-controlled release of small molecules with a rotaxane actuator.

Force-controlled release of small molecules offers great promise for the delivery of drugs and the release of healing or reporting agents in a medical or materials context1-3. In polymer mechanochemistry, polymers are used as actuators to stretch mechanosensitive molecules (mechanophores)4. This technique has enabled the release of molecular cargo by rearrangement, as a direct5,6 or indirect7-10 consequence of bond scission in a mechanophore, or by dissociation of cage11, supramolecular12 or metal complexes13,14, and even by 'flex activation'15,16. However, the systems described so far are limited in the diversity and/or quantity of the molecules released per stretching event1,2. This is due to the difficulty in iteratively activating scissile mechanophores, as the actuating polymers will dissociate after the first activation. Physical encapsulation strategies can be used to deliver a larger cargo load, but these are often subject to non-specific (that is, non-mechanical) release3. Here we show that a rotaxane (an interlocked molecule in which a macrocycle is trapped on a stoppered axle) acts as an efficient actuator to trigger the release of cargo molecules appended to its axle. The release of up to five cargo molecules per rotaxane actuator was demonstrated in solution, by ultrasonication, and in bulk, by compression, achieving a release efficiency of up to 71% and 30%, respectively, which places this rotaxane device among the most efficient release systems achieved so far1. We also demonstrate the release of three representative functional molecules (a drug, a fluorescent tag and an organocatalyst), and we anticipate that a large variety of cargo molecules could be released with this device. This rotaxane actuator provides a versatile platform for various force-controlled release applications.

COVID-19 vaccination affects short-term anti-coagulation levels in warfarin treatment.

Vaccines against SARS-CoV-2 have been recommended across the world, yet no study has investigated whether COVID-19 vaccination influences short-term warfarin anti-coagulation levels. Patients on stable warfarin treatment who received anti-SARS-CoV-2 vaccination were prospectively enrolled and followed up for three months. INR values less than 10 days before vaccination (baseline), 3-5 days (short-term) and 6-14 days (medium-term) after vaccination were recorded as INR0, INR1, and INR2, respectively. The variations of INR values within individuals were compared, and the linear mixed effect model was used to evaluate the variations of INR values at different time points. Logistic regression analysis was performed to determine covariates related to INR variations after COVID-19 vaccination. Vaccination safety was also monitored. There was a significant difference in INR values between INR0 and INR1 (2.15 vs. 2.26, p = 0.003), yet no marked difference was found between INR0 and INR2. The linear mixed effect model also demonstrated that INR variation was significant in short-term but not in medium-term or long-term period after vaccination. Logistic regression analysis showed that no investigated covariates, including age, vaccine dose, genetic polymorphisms of VKORC1 and CYP2C9 etc., were associated with short-term INR variations. Two patients (2.11%) reported gingival hemorrhage in the short-term due to increased INR values. The overall safety of COVID-19 vaccines for patients on warfarin was satisfying. COVID-19 vaccines may significantly influence warfarin anticoagulation levels 3-5 days after vaccination. We recommend patients on warfarin to perform at least one INR monitoring within the first week after COVID-19 vaccination.

Quantitation of levetiracetam concentrations in plasma and saliva samples by ultra-performance liquid chromatography-tandem mass spectrometry: Application to therapeutic drug monitoring for pregnant women with epilepsy.

Although levetiracetam (LEV) has favorable linear pharmacokinetic properties, therapeutic drug monitoring (TDM) is necessary for pregnant women with epilepsy. This study aims to build a simple, reliable, and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determining LEV concentrations in plasma and saliva samples, to support the routine TDM of LEV in Chinese pregnant women with epilepsy. The stable isotope-labeled LEV-d6 was used as the internal standard. The extracted samples were analyzed using a UPLC-MS/MS system with positive electrospray ionization. Mobile phase A was water containing 5 mM ammonium acetate and 0.1% formic acid, and phase B was 1:1 methanol-acetonitrile with 0.1% formic acid. The method was validated and utilized to determine LEV concentrations in non-pregnant and pregnant patients with epilepsy. The developed method was validated in both plasma and saliva samples over a concentration range of 0.1-50 µg/mL. The intra- and inter-batch accuracy for LEV ranged from -7.0% to 2.9%, with precisions between 2.7% and 9.3%. In pregnant patients, the mean dose-standardized LEV trough plasma concentrations were significantly lower than those in non-pregnant patients (4.73 ± 2.99 vs. 7.74 ± 3.59 ng/mL per mg/day; P < 0.0001). It is recommended that the TDM of LEV should be routinely performed during the different stages of pregnancy.

Mechanism insights into the histopathological changes of polypropylene microplastics induced gut and liver in zebrafish.

Microplastics (MPs), emerging as significant pollutants, have been consistently detected in aquatic environments, with the Yangtze River experiencing a particularly severe level of microplastic pollution, exceeding all other watersheds in China. Polypropylene (PP), the plastic most abundantly found in the middle and lower reaches of the Yangtze River Basin, has less comprehensive research results into its toxic effects. Consequently, the present investigation employed zebrafish as a model organism to delve into the toxicological impacts of polypropylene microplastics (PP-MPs) with a diameter of 5 µm across varying concentrations (300 mg/L and 600 mg/L). Using histopathological, microbiota profiling, and transcriptomic approaches, we systematically evaluated the impact of PP-MPs exposure on the intestine and liver of zebrafish. Histopathological analysis revealed that exposure to PP-MPs resulted in thinner intestinal walls, damaged intestinal mucosa, and hepatic cellular damage. Intestinal microbiota profiling demonstrated that, the richness, uniformity, diversity, and homogeneity of gut microbes significantly increased after the PP-MPs exposure at high concentration. These alterations were accompanied by shifts in the relative abundance of microbiota associated with intestinal pathologies, suggesting a profound impact on the intestinal microbial community structure. Concurrently, hepatic transcriptome analysis and RT-qPCR indicated that the downregulation of pathways and genes associated with cell proliferation regulation and DNA damage repair mechanisms contributed to hepatic cellular damage, ultimately exerting adverse effects on the liver. Correlation analysis between the intestinal microbiota and liver transcriptome profiles further highlighted significant associations between intestinal microbiota and the downregulated hepatic pathways. Collectively, these results provide novel insights into the subacute toxicological mechanisms of PP-MPs in aquatic organisms and highlight the need for further research on the ecological and health risks associated with PP-MPs pollution.

Progress of ligand-modified agarose microspheres for protein isolation and purification.

Proteins are the material basis of life and the primary carriers of life activities, containing various impurities that must be removed before use. To keep pace with the increasing complexity of protein samples, it is essential to constantly work on developing new purification technologies for downstream processes. While traditional downstream purification methods rely heavily on protein A affinity chromatography, there is still a lot of interest in finding safer and more cost-effective alternatives to protein A. Many non-affinity ligands and technologies have also been developed in biological purification recently. Here, the current status of biotechnology and the progress of protein separation technology from 2018 to 2023 are reviewed from the aspects of new preparation methods and new composite materials of commonly used separation media. The research status of new ligands with different mechanisms of action was reviewed, including the expanded application of affinity ligands, the development prospect of biotechnology such as polymer grafting, continuous column technology, and its new applications.

Conjugated Polymer Nanoparticles for Tumor Theranostics.

Water-dispersible conjugated polymer nanoparticles (CPNs) have demonstrated great capabilities in biological applications, such as in vitro cell/subcellular imaging and biosensing, or in vivo tissue imaging and disease treatment. In this review, we summarized the recent advances of CPNs used for tumor imaging and treatment during the past five years. CPNs with different structures, which have been applied to in vivo solid tumor imaging (fluorescence, photoacoustic, and dual-modal) and treatment (phototherapy, drug carriers, and synergistic therapy), are discussed in detail. We also demonstrated the potential of CPNs as cancer theranostic nanoplatforms. Finally, we discussed current challenges and outlooks in this field.

Restructuring the Interface of Silk-Polycaprolactone Biocomposites Using Rigid-Flexible Agents.

Natural fiber-reinforced biocomposites with excellent mechanical and biological properties have attractive prospects for internal medical devices. However, poor interfacial adhesion between natural silk fiber and the polymer matrix has been a disturbing issue for such applications. Herein, rigid-flexible agents, such as polydopamine (PDA) and epoxy soybean oil (ESO), were introduced to enhance the interfacial adhesion between Antheraea pernyi (Ap) silk and a common medical polymer, polycaprolactone (PCL). We compared two strategies of depositing PDA first (Ap-PDA-ESO) and grafting ESO first (Ap-ESO-PDA). The rigid-flexible interfacial agents introduced multiple molecular interactions at the silk-PCL interface. The "Ap-PDA-ESO" strategy exhibited a greater enhancement in interfacial adhesion, and interfacial toughening mechanisms were proposed. This work sheds light on engineering strong and tough silk fiber-based biocomposites for biomedical applications.

Efficacy, mechanism, and safety of melatonin-loaded on thermosensitive nanogels for rabbit VX2 tumor embolization: A novel design.

Transarterial chemoembolization (TACE) has been widely used for hepatocellular carcinoma. Reducing hypoxia in the tumor microenvironment after TACE remains a challenge as tumor progression is common in post-TACE patients due to the hypoxic tumor microenvironment. In this study, melatonin loaded on p(N-isopropyl-acrylamide-co-butyl methylacrylate) (PIB-M) was used for tumor embolism. Two types of human hepatoma cell lines were used to explore the mechanism by which melatonin prevents the growth and metastasis of cancer cells in vitro. A VX2 rabbit tumor model was used to evaluate the efficacy, mechanism, and safety of PIB-M in vivo. We found that under hypoxic condition, melatonin could inhibit tumor cell proliferation and migration by targeting hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor A (VEGF-A) in vitro. In vivo, PIB-M inhibited tumor growth and metastasis in rabbit VX2 tumors by promoting apoptosis of tumor cells and targeting related angiogenic proteins and vascular permeability proteins. A high concentration of melatonin in the PIB-M group could be maintained in tumor tissue for 72 h after embolization. The liver and kidney functions were most damaged on the first day but recovered to normal on the seventh day after embolization in the PIB-M group. This novel method may open avenues for reduction of tumor growth and metastasis after TACE and is efficacy and safety, which may be used for treatment for other solid tumors and clinical translation.

Poly(acrylic acid)-Assisted Intrafibrillar Mineralization of Type I Collagen: A Review.

The mineralization of type I collagen is a biological process occurring in vertebrates by which some hard tissues such as bone and dentin are constructed. Due to the extensive clinical needs for bone defect repair and remineralization of mineral-depleted dentin, biomimetic mineralization of collagen is attracting more and more interests. Synthetic analogs of noncollagenous proteins are necessary for directing the in vitro mineralization. In this paper, the function and mechanism of poly(acrylic acid) (PAA) in regulating the mineralization, especially intrafibrillar mineralization (IM) of collagen are reviewed. As two mineralization patterns (extrafibrillar and intrafibrillar) co-exist in natural hard tissues, differences between them in terms of microstructure, biodegradation, cytocompatibility, osteoinduction in vitro, and performance in vivo are systematically compared. Then the roles of PAA in biomimetic collagen IM within one-analog and two-analog systems are discussed, respectively. Moreover, mineralization of some self-mineralizable collagen matrices is described. Due to the interactions between collagen and PAA play a crucial role in the processes of collagen mineralization, some reference researches are also provided involving the collagen/PAA interactions in some other fields. Finally, this review is ended with an outlook for future potential improvements based on the collection of existing bottlenecks in this field.

LRP6/filamentous-actin signaling facilitates osteogenic commitment in mechanically induced periodontal ligament stem cells.

BACKGROUND: Mechanotransduction mechanisms whereby periodontal ligament stem cells (PDLSCs) translate mechanical stress into biochemical signals and thereby trigger osteogenic programs necessary for alveolar bone remodeling are being deciphered. Low-density lipoprotein receptor-related protein 6 (LRP6), a Wnt transmembrane receptor, has been qualified as a key monitor for mechanical cues. However, the role of LRP6 in the mechanotransduction of mechanically induced PDLSCs remains obscure. METHODS: The Tension System and tooth movement model were established to determine the expression profile of LRP6. The loss-of-function assay was used to investigate the role of LRP6 on force-regulated osteogenic commitment in PDLSCs. The ability of osteogenic differentiation and proliferation was estimated by alkaline phosphatase (ALP) staining, ALP activity assay, western blotting, quantitative real-time PCR (qRT-PCR), and immunofluorescence. Crystalline violet staining was used to visualize cell morphological change. Western blotting, qRT-PCR, and phalloidin staining were adopted to affirm filamentous actin (F-actin) alteration. YAP nucleoplasmic localization was assessed by immunofluorescence and western blotting. YAP transcriptional response was evaluated by qRT-PCR. Cytochalasin D was used to determine the effects of F-actin on osteogenic commitment and YAP switch behavior in mechanically induced PDLSCs. RESULTS: LRP6 was robustly activated in mechanically induced PDLSCs and PDL tissues. LRP6 deficiency impeded force-dependent osteogenic differentiation and proliferation in PDLSCs. Intriguingly, LRP6 loss caused cell morphological aberration, F-actin dynamics disruption, YAP nucleoplasmic relocation, and subsequent YAP inactivation. Moreover, disrupted F-actin dynamics inhibited osteogenic differentiation, proliferation, YAP nuclear translocation, and YAP activation in mechanically induced PDLSCs. CONCLUSIONS: We identified that LRP6 in PDLSCs acted as the mechanosensor regulating mechanical stress-inducible osteogenic commitment via the F-actin/YAP cascade. Targeting LRP6 for controlling alveolar bone remodeling may be a prospective therapy to attenuate relapse of orthodontic treatment.

One-Abutment at One-Time in Posterior Edentulism: A Systematic Review and Meta-Analysis.

OBJECTIVE: The objective of this systematic review and meta-analysis was to investigate the clinical significance of one-abutment at one-time protocol in healed posterior edentulism. METHODS: An online search was undertaken in November 2022, which included PubMed, Cochrane Library, Wiley Online Library, and Google Scholar in addition to manual searching. The Cochrane Collaboration tool was performed to assess the quality of selected articles. Marginal bone loss (MBL) was estimated by the performance of meta-analysis. Moreover, all the pooled analyses were based on random-effect models. Subgroup analysis was applied to evaluate the effects of different variables. RESULTS: In line with the inclusion criteria, 6 trials with 446 dental implants were identified. The meta-analysis showed a total of 0.22 mm less MBL within 6 months and decreased by 0.30 mm at 1-year follow-up in favor of one-abutment at one-time protocol. A significant loss MBL was found in implants placed equicrestally using one-abutment at one-time protocol [6 months: mean difference (MD): -0.22 mm; 95% CI, -0.34 to 0.10 mm, P =0.0004; 12 months: MD: -0.32 mm; 95% CI, -0.40 to -0.24 mm, P <0.00001), whereas no difference was found between 2 groups in an implant placed subscrestally (6 months: MD: 0.14 mm; 95% CI, -0.03 to 0.22 mm; P =0.11; 12 months: MD: -0.12 mm; 95% CI, -0.32 to 0.08 mm; P =0.23). CONCLUSIONS: Implant platform position might greatly affect the marginal bone level. Moreover, one-abutment at one-time protocol demonstrated better bone preservation in implants placed equicrestally in healed posterior edentulism. CLINICAL RELEVANCE: This study highlights the significant clinical application of one-abutment at one-time protocol in healed posterior edentulism.

Recent Advances in Bioengineering Bone Revascularization Based on Composite Materials Comprising Hydroxyapatite.

The natural healing process of bone is impaired in the presence of tumors, trauma, or inflammation, necessitating external assistance for bone regeneration. The limitations of autologous/allogeneic bone grafting are still being discovered as research progresses. Bone tissue engineering (BTE) is now a crucial component of treating bone injuries and actively works to promote vascularization, a crucial stage in bone repair. A biomaterial with hydroxyapatite (HA), which resembles the mineral makeup of invertebrate bones and teeth, has demonstrated high osteoconductivity, bioactivity, and biocompatibility. However, due to its brittleness and porosity, which restrict its application, scientists have been prompted to explore ways to improve its properties by mixing it with other materials, modifying its structural composition, improving fabrication techniques and growth factor loading, and co-cultivating bone regrowth cells to stimulate vascularization. This review scrutinizes the latest five-year research on HA composite studies aimed at amplifying vascularization in bone regeneration.

Increased bio-toxicity of leachates from polyvinyl chloride microplastics during the photo-aging process in the presence of dissolved organic matter.

The pollution caused by microplastics (MPs) has gained global attention due to their potential risks to organisms and human health. The process of photo-aging, which plays a crucial role in the transformation of MPs in aquatic environments, has the potential to influence the ecological risk posed by these particles. Dissolved organic matter (DOM) is a prevalent photosensitizer in surface waters that has been shown to facilitate the transformation of various organic compounds by generating reactive oxygen species under light irradiation. The present study investigated the influence of humic acid (HA), a typical component of DOM, on the photo-aging process of polyvinyl chloride MPs (PVC-MPs), using Fourier transform infrared spectroscopy, as well as assessing the resulting ecological risk through bioassays. The results revealed that the presence of HA enhanced the photo-aging of PVC-MP. Moreover, the leachate exhibited higher acute and genetic toxicity under light irradiation when compared to dark conditions. Notably, the presence of HA significantly increased the toxicity of the leachate, emphasizing the need to consider the impact of DOM when assessing the ecological risk of MPs in surface waters. These findings contribute to a more comprehensive understanding of the potential risks associated with microplastic pollution in natural environments.

AAV1.NT-3 gene therapy for X-linked Charcot-Marie-Tooth neuropathy type 1.

X-linked Charcot-Marie-Tooth neuropathy (CMTX) is caused by mutations in the gene encoding Gap Junction Protein Beta-1 (GJB1)/Connexin32 (Cx32) in Schwann cells. Neurotrophin-3 (NT-3) is an important autocrine factor supporting Schwann cell survival and differentiation and stimulating axon regeneration and myelination. Improvements in these parameters have been shown previously in a CMT1 model, TremblerJ mouse, with NT-3 gene transfer therapy. For this study, scAAV1.tMCK.NT-3 was delivered to the gastrocnemius muscle of 3-month-old Cx32 knockout (KO) mice. Measurable levels of NT-3 were found in the serum at 6-month post gene delivery. The outcome measures included functional, electrophysiological and histological assessments. At 9-months of age, NT-3 treated mice showed no functional decline with normalized compound muscle action potential amplitudes. Myelin thickness and nerve conduction velocity significantly improved compared with untreated cohort. A normalization toward age-matched wildtype histopathological parameters included increased number of Schmidt-Lanterman incisures, and muscle fiber diameter. Collectively, these findings suggest a translational application to CMTX1.

In vitro and in vivo evaluation of DC-targeting PLGA nanoparticles encapsulating heparanase CD4+ and CD8+ T-cell epitopes for cancer immunotherapy.

Heparanase has been identified as a universal tumor-associated antigen, but heparanase epitope peptides are difficult to recognize. Therefore, it is necessary to explore novel strategies to ensure efficient delivery to antigen-presenting cells. Here, we established a novel immunotherapy model targeting antigens to dendritic cell (DC) receptors using a combination of heparanase CD4+ and CD8+ T-cell epitope peptides to achieve an efficient cytotoxic T-cell response, which was associated with strong activation of DCs. First, pegylated poly(lactic-coglycolic acid) (PLGA) nanoparticles (NPs) were used to encapsulate a combined heparanase CD4+ and CD8+ T-cell epitope alone or in combination with Toll-like receptor 3 and 7 ligands as a model antigen to enhance immunogenicity. The ligands were then targeted to DC cell-surface molecules using a DEC-205 antibody. The binding and internalization of these PLGA NPs and the activation of DCs, the T-cell response and the tumor-killing effect were assessed. The results showed that PLGA NPs encapsulating epitope peptides (mHpa399 + mHpa519) could be targeted to and internalized by DCs more efficiently, stimulating higher levels of IL-12 production, T-cell proliferation and IFN-γ production by T cells in vitro. Moreover, vaccination with DEC-205-targeted PLGA NPs encapsulating combined epitope peptides exhibited higher tumor-killing efficacy both in vitro and in vivo. In conclusion, delivery of PLGA NP vaccines targeting DEC-205 based on heparanase CD4+ and CD8+ T-cell epitopes are suitable immunogens for antitumor immunotherapy and have promising potential for clinical applications.

Application and Development of Shape Memory Micro/Nano Patterns.

Shape memory polymers (SMPs) are a class of smart materials that change shape when stimulated by environmental stimuli. Different from the shape memory effect at the macro level, the introduction of micro-patterning technology into SMPs strengthens the exploration of the shape memory effect at the micro/nano level. The emergence of shape memory micro/nano patterns provides a new direction for the future development of smart polymers, and their applications in the fields of biomedicine/textile/micro-optics/adhesives show huge potential. In this review, the authors introduce the types of shape memory micro/nano patterns, summarize the preparation methods, then explore the imminent and potential applications in various fields. In the end, their shortcomings and future development direction are also proposed.

Recombinase polymerase amplification assay for rapid detection of Seneca Valley Virus.

Seneca Valley virus (SVV) is related to vesicular disease in pigs, and its clinical symptoms are indistinguishable from other notifiable clinical symptoms of vesicular disease such as foot-and-mouth disease. The rapid and accurate detection of SVV is essential to confirm the pathogenic factors and initiate the implementation of control measures. The development of a rapid, simple, convenient, and low-cost molecular (nucleic acid amplification) test that can be used at the sample collection point has been identified as a key component for controlling SVV. This study describes the development and demonstration of recombinase polymerase amplification (RPA) test targeting the conserved regions of SVV for detection of SVV. The Primers and probes designed by us have shown good sensitivity and specificity in RPA test, which is helpful for RPA to be an effective tool for rapid diagnosis of SVV.

Plaque control alleviated renal damage that was aggravated by experimental periodontitis in obese rats.

OBJECTIVE: This study aimed to evaluate the influence of experimental periodontitis on renal damage in obese rats. MATERIALS AND METHODS: Thirty-two male Sprague Dawley rats were randomly allocated into 4 groups with 8 animals each: obese rats (obese group), obese rats with periodontitis (periodontitis obese group), obese rats with periodontitis that underwent plaque control (plaque-control obese group), and healthy rats (healthy group). Rats were fed a high-fat diet to establish an obesity model. Experimental periodontitis was induced by local ligation with silk around the bilateral maxillary second molars. The plaque control was accomplished by removing ligations and local wiping with an antiseptic rinse. Histology was used to observe the gingival inflammation and clinical attachment level (CAL) to further assess bone loss and to also observe renal structure. Serum creatinine, urea nitrogen, and kidney injury molecule-1 (KIM-1) levels were measured to evaluate renal function. Renal Toll-like receptor 4 (TLR4), nuclear factor-kappa B (NF-κB), serum C-reactive protein (CRP), lipopolysaccharides (LPS), and interleukin-1ß (IL-1ß) were measured to evaluate renal and systemic inflammation. RESULTS: Periodontal histology showed that in the periodontitis obese group, the epithelial barrier was considerably eroded by inflammatory cells, which infiltrated into the subepithelial connective tissue and lamina propria. A periodontal pocket was forming accompanied by the loss of attachment. The extent of infiltration of inflammatory cells and the CAL were significantly higher than those of the obese group (p < .001). In the plaque-control obese group, although the inflammatory condition was significantly improved than in the periodontitis obese group, the clinical attachment level with the presence of fiber hyperplasia could not be restored. Renal histology showed that renal tubular structural damage was aggravated in the periodontitis obese group, including vacuolar degeneration, exfoliation of the proximal tubular epithelial cell lining, multifocal loss of the brush border, and movement of several nuclei from the basement membrane to the lumen. These alterations were improved in the plaque-control obese group. Kidney TLR4 and NF-κB mRNA levels increased significantly in the periodontitis obese group compared to the obese group (p = .015 and p = .015, respectively) and decreased significantly in the plaque-control obese group (p = .028 and p = .021, respectively). Kidney TLR4 and NF-κB protein expression in the plaque-control obese group were significantly lower than those in the periodontitis obese group (p < .001 and p = .043, respectively). Serum creatinine and KIM-1 levels significantly decreased in the plaque-control obese group compared to the periodontitis obese group (p = .001 and p = .002, respectively). At 21 weeks (1 week after periodontal ligation), serum CRP levels in the periodontitis obese group were significantly higher than that in the healthy group (p = .017). Other serum inflammatory markers (LPS and IL-1ß) did not change significantly. CONCLUSION: Experimental periodontitis induced dysfunction and structural destruction of the kidney in obese rats. Plaque control relieved renal damage.

Efficacy and safety of PEGylated exenatide injection (PB-119) in treatment-naive type 2 diabetes mellitus patients: a Phase II randomised, double-blind, parallel, placebo-controlled study.

AIMS/HYPOTHESIS: Glucagon-like peptide 1 receptor agonists (GLP-1 RA) such as exenatide are used as monotherapy and add-on therapy for maintaining glycaemic control in patients with type 2 diabetes mellitus. The current study investigated the safety and efficacy of once-weekly PB-119, a PEGylated exenatide injection, in treatment-naive patients with type 2 diabetes. METHODS: In this Phase II, randomised, placebo-controlled, double-blind study, we randomly assigned treatment-naive Chinese patients with type 2 diabetes in a 1:1:1:1 ratio to receive subcutaneous placebo or one of three subcutaneous doses of PB-119 (75, 150, and 200 µg) for 12 weeks. The primary endpoint was the change in HbA1c from baseline to week 12, and other endpoints were fasting plasma glucose, 2 h postprandial glucose (PPG), and proportion of patients with HbA1c < 53 mmol/mol (<7.0%) and ≤48 mmol/mol (≤6.5%) at 2, 4, 8 and 12 weeks of treatment. Safety was assessed in all patients who received at least one dose of study drug. RESULTS: We randomly assigned 251 patients to one of the four treatment groups (n = 62 in placebo and 63 each in PB-119 75 µg, 150 µg and 200 µg groups). At the end of 12 weeks, mean differences in HbA1c in the treatment groups were -7.76 mmol/mol (95% CI -9.23, -4.63, p < 0.001) (-0.72%, 95% CI -1.01, -0.43), -12.89 mmol/mol (95% CI -16.05, -9.72, p < 0.001) (-1.18%, 95% CI -1.47, -0.89) and -11.14 mmol/mol (95% CI -14.19, -7.97, p <0 .001) (-1.02%, 95% CI -1.30, -0.73) in the 75 µg, 150 µg and 200 µg PB-119 groups, respectively, compared with that in the placebo group after adjusting for baseline HbA1c. Similar results were also observed for other efficacy endpoints across different time points. There was no incidence of treatment-emergent serious adverse event, severe hypoglycaemia or death. CONCLUSIONS/INTERPRETATION: All tested PB-119 doses had superior efficacy compared with placebo and were safe and well tolerated over 12 weeks in treatment-naive Chinese patients with type 2 diabetes. TRIAL REGISTRATION: ClinicalTrials.gov NCT03520972 FUNDING: The study was funded by National Major Scientific and Technological Special Project for Significant New Drugs Development and PegBio.

The adenosine A2A receptor alleviates postoperative delirium-like behaviors by restoring blood cerebrospinal barrier permeability in rats.

Postoperative delirium (POD) is a common post-operative complication in elderly patients that is associated with increased morbidity and mortality. However, the neuropathogenesis of this complication remains unknown. The blood-cerebrospinal fluid barrier (BCB) and brain-blood barrier (BBB) are composed of tight junctions between cells that form physical barriers, and BBB damage plays an important role in the neuropathogenesis of POD. Nevertheless, the role of BCB in POD remains to be elucidated. Herein, we investigated the effect of adenosine A2A receptor (A2A R), a key regulator of the permeability of barriers, on surgery-induced increased permeability of BCB and POD-like behaviors. Open field, buried food, and Y maze tests were used to evaluate behavioral changes in rats after surgery. Levels of tight junction proteins, adherens junction proteins, A2A R, GTP-RhoA, and ROCK2 in the choroid plexus were assessed by western blotting. The concentrations of NaFI and FITC-dextran in the cerebrospinal fluid (CSF) were detected by fluorescence spectrophotometry. Transmission electron microscopy was applied to observe the ultrastructure of the choroid plexus. Surgery/anesthesia decreased the levels of tight junction (e.g., ZO-1, occludin, and claudin1) proteins, increased concentrations of NaFI and FITC-dextran in CSF, damaged the ultrastructure of choroid plexus, and induced POD-like behaviors in rats. An A2A R antagonist alleviated POD-like behaviors in rats. Furthermore, the A2A R antagonist increased the levels of tight junction proteins and restored the permeability of BCB in rats with POD. Fasudil, a selective Rho-associated protein kinase 2 (ROCK2) inhibitor, ameliorated POD-like behaviors induced by A2A R activation. Moreover, fasudil also abolished the increased levels of GTP-RhoA/ROCK2, decreased levels of tight junction proteins, and increased permeability of BCB caused by A2A R activation. Our findings demonstrate that A2A R might participate in regulating BCB permeability in rats with POD via the RhoA/ROCK2 signaling pathway, which suggests the potential of A2A R as a therapeutic target for POD.