Novel strategy for Poxviridae prevention: Thermostable combined subunit vaccine patch with intense immune response.

Poxviruses gained international attention due to the sharp rise in monkeypox cases in recent years, highlighting the urgent need for the development of a secure and reliable vaccine. This study involved the development of an innovative combined subunit vaccine (CSV) targeting poxviruses, with lumpy skin disease virus (LSDV) serving as the model virus. To this end, the potential sites for poxvirus vaccines were fully evaluated to develop and purify four recombinant proteins. These proteins were then successfully delivered to the dermis in a mouse model by utilizing dissolvable microneedle patches (DMPs). This approach simplified the vaccination procedure and significantly mitigated the associated risk. CSV-loaded DMPs contained four recombinant proteins and a novel adjuvant, CpG, which allowed DMPs to elicit the same intensity of humoral and cellular immunity as subcutaneous injection. Following immunization with SC and DMP, the mice exhibited notable levels of neutralizing antibodies, albeit at a low concentration. It is noteworthy that the CSV loaded into DMPs remained stable for at least 4 months at room temperature, effectively addressing the storage and transportation challenges. Based on the study findings, CSV-loaded DMPs are expected to be utilized worldwide as an innovative technique for poxvirus inoculation, especially in underdeveloped regions. This novel strategy is crucial for the development of future poxvirus vaccines.

A Novel Methacryloyl Chitosan Hydrogel Microneedles Patch with Sustainable Drug Release Property for Effective Treatment of Psoriasis.

Psoriasis is a chronic and recurrent skin disease that often requires long-term treatment, and topical transdermal drug delivery can reduce systemic side effects. However, it is still a challenge in efficient transdermal drug delivery for psoriasis treatment due to low penetration efficiency of most drugs and the abnormal skin conditions of psoriasis patients. Here, a safe and effective methacryloyl chitosan hydrogel microneedles (CSMA hMNs) patch is developed and served as a sustained drug release platform for the treatment of psoriasis. By systematically optimizing the CSMA preparation, CSMA hMNs with excellent morphological characteristics and strong mechanical properties (0.7 N needle-1 ) are prepared with a concentration of only 3% (w/v) CSMA. As a proof-of-concept, methotrexate (MTX) and nicotinamide (NIC) are loaded into CSMA hMNs patch, which can produce a sustained drug release of 80% within 24 h in vitro. In vivo experiments demonstrated that the CSMA hMNs patch can effectively inhibit the skin thickening and spleen enlargement of psoriatic mice and has a good biosafety profile at sufficient therapeutic doses. This study provides a new idea for the preparation of hMN systems using modified CS or other biocompatible materials and offers an effective therapeutic option for psoriasis treatment.

Using A Protoplast Transformation System to Enable Functional Studies in Mangifera indica L.

Mangoes (Mangifera indica L.) are an important kind of perennial fruit tree, but their biochemical testing method and transformation technology were insufficient and had not been rigorously explored. The protoplast technology is an excellent method for creating a rapid and effective tool for transient expression and transformation assays, particularly in plants that lack an Agrobacterium-mediated plant transformation system. This study optimized the conditions of the protoplast isolation and transformation system, which can provide a lot of help in the gene expression regulation study of mango. The most beneficial protoplast isolation conditions were 150 mg/mL of cellulase R-10 and 180 mg/mL of macerozyme R-10 in the digestion solution at pH 5.6 and 12 h of digestion time. The 0.16 M and 0.08 M mannitol in wash solution (WI) and suspension for counting (MMG), respectively, were optimal for the protoplast isolation yield. The isolated leaf protoplasts (~5.4 × 105 cells/10 mL) were transfected for 30 min mediated by 40% calcium-chloride-based polyethylene glycol (PEG)-4000-CaCl2, from which 84.38% of the protoplasts were transformed. About 0.08 M and 0.12 M of mannitol concentration in MMG and transfection solutions, respectively, were optimal for protoplast viability. Under the florescence signal, GFP was seen in the transformed protoplasts. This showed that the target gene was successfully induced into the protoplast and that it can be transcribed and translated. Experimental results in this paper show that our high-efficiency protoplast isolation and PEG-mediated transformation protocols can provide excellent new methods for creating a rapid and effective tool for the molecular mechanism study of mangoes.

An Integrated Therapeutic and Preventive Nanozyme-Based Microneedle for Biofilm-Infected Diabetic Wound Healing.

The healing of biofilm-infected diabetic wounds characterized by a deteriorative tissue microenvironment represents a substantial clinical challenge. Current treatments remain unsatisfactory due to the limited antibiofilm efficacy caused by weak tissue and biofilm permeability of drugs and the risk of reinfection during the healing process. To address these issues, an integrated therapeutic and preventive nanozyme-based microneedle (denoted as Fe2 C/GOx@MNs) is engineered. The dissolvable tips with enough mechanical strength can deliver and rapidly release Fe2 C nanoparticles (NPs)/glucose oxidase (GOx) in the biofilm active regions, enhancing tissue and biofilm permeability of Fe2 C NPs/GOx, ultimately achieving highly efficient biofilm elimination. Meanwhile, the chitosan backing layer can not only act as an excellent physical barrier between the wound bed and the external environment, but also prevent the bacterial reinvasion during wound healing with its superior antibacterial property. Significantly, the biofilm elimination and reinfection prevention abilities of Fe2 C/GOx@MNs on wound healing are proved on methicillin-resistant Staphylococcus aureus-biofilm-infected diabetic mouse model with full-thickness wound. Together, these results demonstrate the promising clinical application of Fe2 C/GOx@MNs in biofilm-infected wound healing.

Personalized demand-responsive biphasic microneedle patch for smart drug administration.

Many patients, especially those with chronic diseases, would benefit from personalized drugs that could modulate the treatment regimen. Tailored drug delivery via microneedle patches (MNPs) has emerged as a promising technology to address this problem. However, it is still difficult to modulate the treatment regimen in one MNP. Here, multiple treatment regimens were achieved by the same MNP functionalized with modifiable nanocontainers (NCs). The MNPs were biphasic in design, resulting in approximately a twice as high drug loading capacity than that of traditional dissolving MNPs. The drug-loaded NCs could have a zero-order release rate for at least 20 d in vitro. Furthermore, three model MNPs, Type-A (100% drug), Type-B (50% drug and 50% NCs) and Type-C (100% NCs) were generated to simulate various personalized dosing needs. In vivo application of these models could achieve effective therapeutic drug concentrations in the first 12 h and adjusted the duration of effective drug action from 24 h to 96 h and 144 h, respectively, with outstanding biocompatibility. These findings indicate that this device holds significant promise for personalized drug delivery.

The Synergistic Mechanism of Photosynthesis and Antioxidant Metabolism between the Green and White Tissues of Ananas comosus var. bracteatus Chimeric Leaves.

Ananas comosus var. bracteatus (Ac. bracteatus) is a typical leaf-chimeric ornamental plant. The chimeric leaves are composed of central green photosynthetic tissue (GT) and marginal albino tissue (AT). The mosaic existence of GT and AT makes the chimeric leaves an ideal material for the study of the synergistic mechanism of photosynthesis and antioxidant metabolism. The daily changes in net photosynthetic rate (NPR) and stomatal conductance (SCT) of the leaves indicated the typical crassulacean acid metabolism (CAM) characteristic of Ac. bracteatus. Both the GT and AT of chimeric leaves fixed CO2 during the night and released CO2 from malic acid for photosynthesis during the daytime. The malic acid content and NADPH-ME activity of the AT during the night was significantly higher than that of GT, which suggests that the AT may work as a CO2 pool to store CO2 during the night and supply CO2 for photosynthesis in the GT during the daytime. Furthermore, the soluble sugar content (SSC) in the AT was significantly lower than that of GT, while the starch content (SC) of the AT was apparently higher than that of GT, indicating that AT was inefficient in photosynthesis but may function as a photosynthate sink to help the GT maintain high photosynthesis activity. Additionally, the AT maintained peroxide balance by enhancing the non-enzymatic antioxidant system and antioxidant enzyme system to avoid antioxidant damage. The enzyme activities of reductive ascorbic acid (AsA) and the glutathione (GSH) cycle (except DHAR) and superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were enhanced, apparently to make the AT grow normally. This study indicates that, although the AT of the chimeric leaves was inefficient at photosynthesis because of the lack of chlorophyll, it can cooperate with the GT by working as a CO2 supplier and photosynthate store to enhance the photosynthetic ability of GT to help chimeric plants grow well. Additionally, the AT can avoid peroxide damage caused by the lack of chlorophyll by enhancing the activity of the antioxidant system. The AT plays an active role in the normal growth of the chimeric leaves.

Do premature and postterm birth increase the risk of epilepsy? An updated meta-analysis.

BACKGROUND: Many studies have reported that premature birth is associated with a higher incidence of epilepsy, and postterm birth also increases the risk of epilepsy. The effects of different gestational ages (GAs) on epilepsy have become a research hotspot, but the findings of these studies remain controversial, and no systematic review has been performed until now. OBJECTIVE: The aim of this study was to evaluate the impact of different GAs on the incidence of epilepsy. DATA SOURCES: The main databases, including PubMed, Medline, Embase, Cochrane Library, and Web of Science, were searched using the terms "preterm/premature/early/postterm/postmature/late/delayed delivery/birth", "gestational age", and "epilepsy/seizure" for eligible studies published up to April 1, 2019. The search was limited to English-language articles. STUDY SELECTION: Observational studies investigating the association between epilepsy and premature or postterm birth were included in this meta-analysis. We only selected studies that had clearly reported GA and the occurrence of epilepsy. DATA EXTRACTION AND ANALYSIS: Two reviewers independently extracted the data. The quality of the included studies was examined in accordance with the Newcastle-Ottawa criteria, and the heterogeneity and publication bias were tested. We used sensitivity and subgroup analyses to determine the source of heterogeneity. A logistic randomized-effects model was used to assess the collected data when I2 ≥ 50%. MAIN OUTCOMES: The primary outcome was the odds ratio (OR) of epilepsy. RESULTS: The research included eleven eligible studies with a total of 4,513,577 participants. Studies involving premature birth showed that the risk of epilepsy was 2.16 times higher in the premature birth group (<37 weeks) than in the full-term birth group (≥37 weeks) (OR [99% confidence interval [CI]] = 2.16 [1.80, 2.58]; P < 0.001). Those born before 32 weeks were associated with an increased occurrence of epilepsy when compared with those born at 32-36 weeks (OR [99% CI] = 2.73 [1.90, 3.94]; P < 0.001). However, the difference in the incidence of epilepsy between postterm children (41 weeks or more) and full-term children (37-40 weeks) was not statistically significant (OR [99% CI] = 1.05 [0.98, 1.12]; P = 0.067). CONCLUSIONS: Preterm birth was closely associated with a higher risk of epilepsy throughout childhood that persisted into adulthood, and the association became stronger as GA decreased, while there was no significant difference in the risk of developing epilepsy between postterm and full-term offspring.