Tracking endogenous proteins based on RNA editing-mediated genetic code expansion.

Protein labeling approaches are important to study proteins in living cells, and genome editing tools make it possible to tag endogenous proteins to address the concerns associated with overexpression. Here we established RNA editing-mediated noncanonical amino acids (ncAAs) protein tagging (RENAPT) to site-specifically label endogenous proteins with ncAAs in living cells. RENAPT labels protein in a temporary and nonheritable manner and is not restricted by protospacer adjacent motif sequence. Using a fluorescent ncAA or ncAA with a bio-orthogonal reaction handle for subsequent dye labeling, we demonstrated that a variety of endogenous proteins can be imaged at their specific subcellular locations. In addition, two proteins can be tagged individually and simultaneously using two different ncAAs. Furthermore, endogenous ion channels and neuron-specific proteins can be real-time labeled in primary neurons. Thus, RENAPT presents a promising platform with broad applicability for tagging endogenous proteins in living cells to study their localization and functions.

Association of remnant cholesterol with CVD incidence: a general population cohort study in Southwest China.

Background: Emerging evidence has indicated that remnant cholesterol (RC) could predict cardiovascular disease (CVD) incidence. Nevertheless, the relationship between RC and CVD risk, especially within the general Chinese population, remains scarce. Objective: The present research aimed to assess whether RC concentrations and CVD outcomes in general Chinese adults are related. Methods: The Cox proportional hazard model was established to explore the relationship between RC and the outcomes of CVD and CVD subgroups. A restricted cubic spline (RCS) was utilized to investigate the dose-response connection between RC and the risk of CVD outcomes, and the ROC curve was used to calculate the corresponding cutoff values. Moreover, stratified analysis was conducted to investigate the potential effect modification in the association between RC and CVD outcomes. Results: Significant positive associations were found between elevated categorical RC and increased risk of CVD (HR Q4, 1.80; 95% CI 1.15-2.79; P-value = 0.008), atherosclerotic cardiovascular disease (HR Q4, 2.00; 95% CI 1.22-3.27; P-value = 0.007), stroke (HR Q4, 1.66; 95% CI 1.02-2.69; P-value = 0.040), and ischemic stroke (HR Q4, 1.87, 95% CI 1.08-3.25; P-value = 0.034), respectively. Our study suggested that the incidence of CVD outcomes increased when RC levels were above 0.75 mmol/L. Importantly, the CVD risks related to RC were more likely to be those found in subjects aged above 60 years, women, subjects with BMI <24 kg/m2, and subjects with hypertension and unhealthy diet patterns. Conclusions: Aberrant high level of RC is associated with elevated CVD risk, independent of low-density lipoprotein cholesterol (LDL-C). Our data reveal urgent primary prevention for subjects with high RC levels to a low incidence of CVD, especially for the elderly, women, and those with hypertension and unhealthy diet patterns.

Delivery of Transcriptional Factors for Activating Antioxidant Defenses against Inflammatory Bowel Disease.

Oxidative stress caused by the overproduction of reactive oxygen species (ROS) plays an important role in inflammatory bowel disease (IBD). It is well-known that the Nrf2-ARE (antioxidative response element) pathway is important in the regulation mechanism of antioxidant defense. Therefore, Nrf2 activation may be an effective therapeutic strategy for IBD. Here, we reported the development of a nucleus-targeted Nrf2 delivery nanoplatform, termed N/LC, that could accumulate in inflamed colonic epithelium, reduce inflammatory responses, and restore epithelium barriers in a murine model of acute colitis. N/LC nanocomposites could quickly escape from lysosomes, so Nrf2 largely accumulated in the nucleus of colonic cells, activated the Nrf2-ARE signaling pathway, further elevated the expression levels of downstream detoxification and antioxidant genes, and protected cells from oxidative damage. These results suggested that N/LC might be a potential nanoplatform for IBD therapy. The study provided the basis for the biomedical applications of Nrf2-based therapeutics in various diseases.

Dibenzocyclooctyne linked lysine-cyclodextrin for efficient intranucleus delivery of proteins.

Intranuclear protein delivery shows great prospects in broadening the application scope of protein therapy and revolutionizing medicine, however, effective delivery of native proteins into the nucleus of cells remains a great challenge. Herein, we report the supramolecular nanoparticles based on the self-assembly of dibenzocyclooctyne (DIBO) linked lysine-cyclodextrin (DLC) for efficient intranucleus delivery of proteins. Coordination-driven self-assembly of DLCs in aqueous solution enables efficient encapsulation of proteins just by simple mixing, so as to maintain their biological activity in a reliable way. DLC nanoparticles ensure effective intranuclear protein delivery for therapeutic applications and gene regulation. This rationally designed DIBO containing amino acid-cyclodextrin derivative allows the development of a convenient and universal nanoplatform for intranuclear delivery of native proteins.

Comparative Study of Dezocine and Ketorolac Tromethamine in Patient-Controlled Intravenous Analgesia of Laparoscopic Cholecystectomy.

Purpose: This study aimed to observe the application value of dezocine and ketorolac tromethamine in patient-controlled intravenous analgesia (PCIA) of patients undergoing laparoscopic cholecystectomy (LC). Methods: A total of 154 patients who underwent LC surgery in our hospital and received PCIA after surgery from September 2020 to September 2021 were selected, they were divided into group A (n = 77) and group B (n = 77). Group A was given dezocine and group B was given ketorolac tromethamine. The analgesia, sedation, comfort, and adverse reactions of the two groups were closely observed at 4, 8, 12, and 24 h after surgery. Results: At 4, 8, 12, and 24 h after surgery, the visual analog scale scores in group B were lower than those in group A (P < 0.05). At 4, 8, 12, and 24 h after surgery, the Ramsay scores in group B were higher than those in group A (P < 0.05). At 4, 8, 12, and 24 h after surgery, there was no significant difference in Bruggrmann comfort scale scores between the two groups (P > 0.05). There was no significant difference in the incidence of adverse reactions between the two groups (P > 0.05). Conclusion: Both dezocine and ketorolac tromethamine have high clinical application value in patients who underwent LC surgery and received PCIA, with higher patient comfort and fewer adverse reactions. But compared with dezocine, ketorolac tromethamine can achieve better sedative and analgesic effects, which is worthy of clinical promotion.

Clickable amino acid tuned self-assembly of a nucleus-selective multi-component nanoplatform for synergistic cancer therapy.

Nucleus-targeted therapy holds great promise in cancer treatment; however, a lack of effective nucleus-specific delivery significantly limits its application potential. Here, we report a nucleus-targeted synergistic chemo-photodynamic therapy based on the self-assembly of chlorin e6 (Ce6) and doxorubicin (DOX) tuned by clickable dibenzocyclooctyne (DIBO) functionalized lysine (D-K) and subsequent reaction with crosslinkers. The assembled nanodrugs with high loading efficiency and long-term stability show enhanced cellular uptake and accumulation in the nucleus, resulting in greatly improved in vitro and in vivo chemo-photodynamic efficacy. Notably, D-K can promote the rapid self-assembly of Ce6 and DOX in aqueous solution, avoiding the introduction of organic solvents or tedious preparations. In addition, the introduction of the DIBO group can effectively expand the types of self-assembly material and enhance the self-assembly behaviour through a copper-free click reaction. Therefore, we present an effective nucleus-targeted combination drug delivery strategy, which has great potential in the treatment of many diseases.

Oral delivery of antioxidant enzymes for effective treatment of inflammatory disease.

Oral administration of protein is very challenging for therapeutic applications due to its instability and easy degradation in the gastrointestinal tract. Herein, we reported an approach to encapsulate native anti-inflammatory proteins in wind chimes like cyclodextrin (WCC) for efficient oral protein delivery. The amphiphilic WCC can self-assemble into nanoparticles in aqueous solution and achieve superior encapsulation of two antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT) by simply mixing with protein solution, avoiding any extra cumbersome steps that might inactivate protein. WCC nanovehicles can effectively protect enzyme activity and enhance their intracellular delivery. SOD and CAT co-loaded WCC nanoparticles (SC/WCC) can integrate the synergistic effect of SOD and CAT for enhancing the removal of reactive oxygen species (ROS), effectively inhibit the inflammatory response by reducing the secretion of proinflammatory factors and protect cells from ROS-induced oxidative damage. In the mouse colitis model, SC/WCC administered orally was able to efficiently accumulate in the inflamed colon, significantly inhibited the expression of proinflammatory mediators and notably alleviated the symptoms related to colitis. Therefore, we believe that the strategies we described here may provide a convenient and powerful platform for the treatment of other inflammatory diseases.

Bakuchiol from Psoralea corylifolia L. Ameliorates acute kidney injury and improves survival in experimental polymicrobial sepsis.

Bakuchiol (BAK) is a prenylated phenolic mono-terpene extracted from the fruit of Psoralea corylifolia L., which exerts a protective effect on organs. However, whether BAK has a protective effect on sepsis-induced acute kidney injury (S-AKI) is not clear. In our study we have demonstrated for the first time that pretreatment with BAK significantly reduced bacterial load, inflammation and renal oxidative stress in caecal ligation and puncture (CLP)-induced sepsis. Moreover, CLP-induced renal histological damage, mortality and clinical signs were markedly attenuated by BAK. Additionally, BAK inhibited sepsis-induced activation of NF-κB and p38 MAPK signaling in the kidneys. The evidence presented here has confirmed that BAK exerts multifunctional activity in protection against S-AKI. This action of BAK is probably due to the blockade of the NF-κB and p38 MAPK signaling pathways. Our findings offer a novel potential for BAK in protection against sepsis and S-AKI.

Highly uniform self-assembled microspheres from single macromolecule self-recognition for enhanced cancer immunotherapy.

Self-assembled highly uniform microspheres from star-shaped biocompatible polymers have been prepared as a local delivery system for co-loading different immunoagents together and ensuring their release in different stages, so as to realize effective cooperative immunotherapy and minimize systemic side effects.

Stimuli-responsive self-assembled dendrimers for oral protein delivery.

Oral delivery of protein is very challenging for therapeutic applications due to protein instability and degradability. Herein we synthesized benzoboroxole-containing multi-armed poly(ethylene glycol) amphiphilic dendrimers sensitive to both pH and glucose. The multi-branched structure of dendrimer facilitated its self-assembly into micelles in aqueous solution and exhibited good encapsulation of insulin just by simply mixing with insulin solution. The insulin loaded micelles were stable under strong acidic condition and showed glucose-responsive insulin release at physiological pH. The reversible binding between benzoboroxole and glucose can effectively regulate the disassembly of micelles and insulin release rate, which in turn stabilizes blood sugar fluctuations. Utilizing a mouse model of type 1 diabetes, it was demonstrated that these insulin loaded micelles could successfully control the blood glucose levels under a normoglycemic state and avoid the risk of hypoglycemia. Therefore, this smart delivery system shows great potential to significantly improve the effect of oral protein therapy.

Quantitative and high drug loading of self-assembled prodrug with defined molecular structures for effective cancer therapy.

Nanomedicines have made significant progress in the delivery of small molecular drugs, many challenges, however, still remain to be overcome, such as unsatisfactory drug loading, formulation instability, premature drug leakage, and poor blood circulation. Herein, an innovative glutathione (GSH)-sensitive amphiphilic dendritic prodrug with quantitative and high drug loading (>30 wt%) is reported. The multi-armed structure of prepared prodrug can self-assemble into nanoparticles in aqueous solution without the introduction of any organic solvents. The self-assembled prodrug nanoparticle is composed of the following key components: (i) polyethylene glycol (PEG) outer shell ensuring biocompatibility and prolonging blood circulation, (ii) prodrug inner core responding to GSH for triggered release of intact drug, (iii) multi-armed dendritic structure facilitating self-assembly and enhancing drug loading content, (iv) covalent drug conjugation avoiding drug leakage and improving stability, (v) defined chemical structures and quantitative drug loading easy for reproduction. Both in vitro and in vivo results show that these GSH-responsive prodrug nanoparticle exhibits significant inhibition of tumor cell growth, and is promising for efficient and safe chemotherapeutic delivery.

Biocompatible anionic polyelectrolyte for improved liposome based gene transfection.

Cationic liposomes have been widely used as efficient gene carriers. However, the serious cytotoxicity caused by exposed positive charges restricts the further application of those kinds of gene vectors. Thus, it is challenging to develop biocompatiable non-positive charge carriers to achieve high gene transfection efficiencies. Herein, we report a novel design by pasting biocompatible anionic polyelectrolyte, namely alginic acid, hyaluronic acid, pectin and polyglutamic acid, to the positive charge surface of liposome/pDNA complex. Through shielding the positive charges, the new gene carriers show decreased cytotoxicity while still maintaining high transfection efficiency. To be noted, the complex formed by coating polyglutamic acid to the surface of liposome/pDNA greatly enhanced the transfection efficiency in HepG2 cells, and the use of pectin shows increased transfection in MCF-7 cells. Hemolysis assay proved a possible mechanism that when the new gene complex was internalized into cells, as acidity increases, more side chains become hydrophobic, and thus destabilizing the endosomal membrane to accelerate DNA escape. The present results suggest that such anionic polyelectrolyte covered liposome based carrier possess promising application for clinical gene delivery.