Respiratory infectious diseases frequently erupt on a global scale, with RNA viruses, such as SARS-CoV-2, RSV, and influenza viruses, posing challenges to vaccine development due to their high mutation rates. Traditional vaccine development cycles are lengthy and struggle to keep pace with rapidly evolving viruses, whereas messenger RNA (mRNA) vaccines have demonstrated significant advantages due to their short development periods, straightforward production, and low costs. After the outbreak of the COVID-19 pandemic, multiple mRNA vaccines, including Pfizer-BioNTech and Moderna, rapidly received emergency use authorization, validating their feasibility. The Nobel Prize in Physiology or Medicine in 2023 was awarded to Katalin Karikó and Drew Weissman, underscoring the efficacy of mRNA vaccine technology. In 2024, the U.S. Food and Drug Administration (FDA) approval of Moderna's respiratory syncytial virus (RSV) mRNA vaccine marked the immense potential of mRNA technology in vaccine innovation. This review article summarizes the design, clinical research, and future challenges of mRNA vaccines for respiratory viruses, delving into antigen design, mRNA delivery systems, and advancements in vaccines for multiple respiratory viruses, including innovations in self-amplifying mRNA and circular mRNA vaccines. Additionally, the development of combination vaccines is underway, aiming to provide protection against multiple viruses through a single administration. Despite the significant progress in mRNA vaccine development, challenges remain regarding raw material costs, stability, and delivery efficiency. In the future, with technological advancements and the accumulation of clinical experience, the design strategies and delivery systems of mRNA vaccines are expected to be continuously optimized, thereby enhancing their safety and efficacy.

MicroRNAs (miRNAs) are crucial in diverse biological processes. In recent years, extensive research has significantly advanced our understanding of miRNA biogenesis, function, and its implications in various biological processes and diseases. In development, miRNAs precisely regulate gene expression to ensure proper organismal growth. miRNAs serve as essential modulators of cell proliferation and differentiation, thereby determining cell fate. Regarding the regulation of apoptosis, miRNAs play a significant role in controlling programmed cell death. Moreover, in metabolism, miRNAs are involved in modulating various pathways. This review examines the latest advancements in miRNA research, encompassing their biogenesis, functional roles, and involvement in various biological processes and diseases. A specific emphasis is placed on the roles of miRNAs in development, cell proliferation, apoptosis, and metabolic regulation. Additionally, cutting-edge technologies were discussed for the potential therapeutic applications of miRNA-based strategies.

Background: Intracerebral hemorrhage (ICH) remains a devastating neurological disorder with limited therapeutic options. Neural stem cell (NSC)-based therapies have emerged as a potential regenerative approach, yet the molecular mechanisms regulating NSC behavior require further elucidation. The role of miR-21 in NSC differentiation and proliferation during ICH recovery remains unexplored.

Methods: In vitro NSC cultures were analyzed for miR-21 expression dynamics during differentiation via qPCR. Lentiviral overexpression and knockdown of miR-21 were employed to assess its functional impact. The SOX2/LIN28-let-7 pathway was investigated using Western blot, luciferase reporter assays, and immunofluorescence. In vivo, miR-21-overexpressing NSCs were transplanted into a murine ICH model, with neurogenesis evaluated by immunostaining and neurological recovery assessed through behavioral tests (mNSS, rotarod).

Results: miR-21 expression significantly increased during NSC differentiation, correlating with reduced SOX2 levels. Mechanistically, miR-21 directly targeted SOX2, disrupting the SOX2/LIN28-let-7 axis to promote NSC proliferation and lineage commitment. In ICH mice, transplantation of miR-21-overexpressing NSCs enhanced neurogenesis and improved motor coordination and neurological deficits at 28 days post-transplantation.

Conclusions: Our findings identify miR-21 as a critical regulator of NSC plasticity through SOX2/LIN28-let-7 signaling, highlighting its therapeutic potential for enhancing neuroregeneration and functional recovery in ICH. Targeting miR-21 may represent a novel strategy to optimize NSC-based therapies for hemorrhagic stroke.

Although hypertension is a frequently seen chronic condition across the world, its exact cause remains unclear. Animal models are beneficial for clarifying the pathogenic mechanism of hypertension and examining new treatments. An optimal animal model for studies on hypertension must well mimic human-like hemodynamics and pathophysiological structural modification, showing human disease features and complications timely or even ahead of time. A review of the most frequently used hypertensive animal models available, including small and large animals, induced and genetic models, would provide an insight into the appropriate targets to be addressed in the development of different hypertensive animal models. Another focus of the review are the processes of target-organs injury caused by high blood pressure, which mainly influences human health.

The COVID-19 pandemic posed a challenge for clinical management of a new lung disease that was characterized by inflammation, endothelial cell dysfunction, and thrombosis, which occur after the replication phase of infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). There are many laboratory models of active SARS-CoV-2 infection in mice, reflecting an acute lung injury in an otherwise healthy animal, but there is a lack of accurate animal models of the postviral inflammatory phase of the COVID-19 lung reflecting severe disease. The monocrotaline (MCT)–treated rat is a widely used laboratory model of pulmonary hypertension (PH). Not often discussed, however, are the observed changes in inflammation, edema, fibrosis, and microthrombosis in the lung prior to PH. At the cellular level, there is loss of pneumocytes and endotheliopathy, and at the molecular level the MCT rat lung is characterized by a pro-inflammatory cytokine profile, namely elevated interleukin 6, transforming growth factor β and tumor necrosis factor, M1 macrophage phenotype, and dysregulation of the angiotensin converting enzyme (ACE)/ACE2 balance. The systems-level pathophysiology of the MCT-treated rat includes progressive cardiopulmonary dysfunction. The MCT-treated rat clearly differs from the COVID-19 lung in terms of the triggers for pathology, but there are many parallels apparent in both the MCT-treated rat and the COVID-19 lung. The MCT-treated rat lung as a model of the COVID-19 lung may provide an in-depth understanding of the factors that drive the lung to more severe pathology, treatments that benefit lung recovery, or the factors that prove a useful research platform for future emerging respiratory threats of similar pathology.

Background: This study investigated the phenolic profile, antioxidant capacity, antibacterial effect, and antihemolytic activity of nettle leaves from two understudied Algerian species, Urtica pilulifera and Urtica urens.

Methods: Urtica pilulifera and Urtica urens leaves extracts were prepared by maceration using methanol and distilled water respectively. Their phytochemical analysis (total phenolic content, flavonoids, hydrolysable and condensed tannins) was determined. The chemical profle of these extracts was performed using ultra-high-performance liquid chromatography diode array detection tandem mass spectrometry. Antioxidant activity (using the 2,2-diphenyl-1-picrylhydrazyl free radical scavenging method), antibacterial activity (using disk diffusion method), and antihemolytic activity were carried out.

Results: Results reveal high levels of flavonoids, tannins, alkaloids, and terpenoids. Several classes of compounds were identified in the extracts. Phenolic acid and other acids and their derivatives and flavonoids were detected in the extracts, with 4-O-caffeoyl-quinic acid and 5-O-caffeoyl-quinic acid being the main constituents in both extracts. Caffeic acid was also the main constituent present only in U. urens extract. For antioxidant activity of the methanolic extract of U. pilulifera and the aqueous extract of U. urens, the percentage inhibition value (82.76% and 59.06%) and the half-maximum inhibitory concentration (IC₅₀) value (302 and 423 μg/mL), respectively, were obtained. Antibacterial activity of nettle extracts (10 mg/mL) demonstrated the sensitivity of pathogenic strain susceptibility. The strongest antibacterial effect on tested strains was found in the aqueous extract of U. urens against Staphylococcus aureus (21 ± 0.41 mm) compared to the methanolic extract of U. pilulifera (16 ± 0.40 mm). The antihemolytic activity in the methanolic and aqueous extracts was 76.26% and 60.67%, respectively. The methanolic extract exhibited exceptional antihemolytic effect, with an IC₅₀ value of 327 μg/mL, whereas the aqueous extract had an IC₅₀ value of 412 μg/mL.

Conclusions: The study confirms the presence of bioactive substances in the nettle species, including flavonoids and tannins, which possess antioxidant, antibacterial, and antihemolytic properties, and can be processed into food and pharmaceutical products.

Background: Sudden sensorineural hearing loss (SSNHL), often associated with tinnitus, significantly impacts individuals' quality of life. Current treatments, such as free drugs via intravenous or intratympanic (IT) administration of dexamethasone (DEX) and lidocaine, face limitations like low bioavailability and rapid drug clearance. To address these challenges, we developed a local co-delivery system combining DEX microcrystals (DEX MCs) and lidocaine-loaded poly(lactic-co-glycolic acid) (PLGA) non-spherical microparticles (LPNMs) for sustained drug release in the inner ear.

Methods: DEX MCs and LPNMs were prepared using the traditional precipitation technique and double emulsion-solvent evaporation, respectively. After characterizing physicochemical properties and drug release kinetics, they were dispersed in sodium hyaluronate solution for IT injection, then in vivo pharmacokinetics and biocompatibility in guinea pigs were studied.

Results: DEX MCs exhibited stable dissolution, while LPNMs provided sustained lidocaine release, reducing potential side effects. In vivo studies in guinea pigs demonstrated prolonged drug retention in the perilymph and improved pharmacokinetics. Histological evaluation confirmed the good biocompatibility of this combined delivery system, with no significant inner ear damage observed.

Conclusion: This co-delivery system can be used as a depot for delivering both DEX and lidocaine to the inner ear and offers a promising approach for the synergistic treatment of SSNHL associated with tinnitus.

Background: Most mutations in the COL6A3 gene lead to collagen VI-related myopathies. This is due to a reduced expression or mislocalization of the COL6A3 protein. Therefore, studying the consequence of knocking out the Col6a3 gene in mouse models is relevant, but the Col6a3 mouse models reported so far do not entirely abolish COL6A3 protein expression.

Methods: Here, we present the development, validation and preliminary phenotypic characterization of a novel CRISPR-based knockout mouse model targeting Col6a3 exon 3 (Col6a3^d3/d3).

Results: In this mouse model, Col6a3 mRNA is still expressed at a similar level to wild-type littermates, although the expected protein is undetectable by mass spectrometry. Histological analysis of Col6a3^d3/d3 quadriceps revealed an abnormally high frequency of muscle cells with internally nucleated muscle cells, consistent with a myopathy phenotype. Interestingly, Col6a3^d3/d3 mice are smaller in size, with their fat, muscle, and bone kept proportional compared to wild-type littermates.

Conclusions: In summary, we performed the validation and preliminary phenotypic characterization of a novel Col6a3 knockout mouse model that could be further characterized and used to study COL6A3 biology and model collagen VI-associated diseases.

Background: Maternal viral infection during pregnancy can lead to maternal immune activation (MIA), increasing the risk of neurodevelopmental disorders in offspring. Amantadine (AMA) exhibits antiviral activity and is widely employed in the management of neurologic conditions. Nevertheless, the efficacy of AMA in treating MIA is currently not established.

Methods: MIA was induced by polyinosinic acid–polycytidylic acid (poly(I:C)); AMA was administered from embryonic (E) day 11.5 for 3 days. BV-2 cells were stimulated using poly(I:C) and treated with AMA. Behavior was assessed via open field test, elevated plus maze test, three-chamber sociability test, and marble burying test. Neuronal morphology was vizualized using Nissl stain; apoptosis via TUNEL (terminal deoxynucleotidyl transferase dUTP nick-end labeling) stain; protein expression (Iba1, NeuN, CD68, TNF-α [tumor necrosis factor-alpha], IL-1β [interleukin-1β]) using immunofluorescence (IF); interleukin-6 (IL-6) levels using enzyme-linked immunosorbent assay; reactive oxygen species using staining; Iba1, NeuN, Bcl-2, Bax, and cleaved caspase 3 using Western blot; and gene expression changes using RNA-seq.

Results: AMA treatment reduced the levels of IL-6 in maternal blood, improved autism-like behaviors in MIA offspring, and effectively prevented neuronal damage and neuroinflammation. In vitro cellular studies have demonstrated that AMA effectively downregulates the expression levels of pro-inflammatory cytokines, including IL-6, TNF-α, and IL-1β. RNA-seq analysis indicated that AMA mitigates abnormal activation of microglia by modulating inflammatory pathways associated with IL-6.

Conclusion: AMA can prevent the development of neuropsychiatric disorders in MIA offspring. This effect may be related to its ability to attenuate neuronal damage, reduce neuronal apoptosis, and inhibit neuroinflammation, indicating that the antiviral drug AMA may be a potential treatment for MIA.

Background: Chaihu Guizhi Decoction (CGD) is a classical prescription in traditional Chinese medicine (TCM) that has been shown to be effective against infection and depression. However, the mechanisms of action of CGD in treating patients with depression who are also infected with a virus remain unknown. This study investigated the pharmacological mechanisms of CGD in treating depression-like mice infected with influenza A virus (IAV).

Methods: The acute depression-like mouse model infected with IAV was established. The effects of CGD on alleviating depression-like symptoms were observed using the sucrose preference test and monoamine neurotransmitter expression. Additionally, the effectiveness of CGD against infection was assessed by examining the relative expression of the M gene and pathological changes. To evaluate the anti-inflammatory effects of CGD, the levels of cytokines were measured, and the Th17/Treg ratio was analyzed to evaluate the immunomodulatory effects of CGD.

Results: The study results show that CGD enhanced the rates of sucrose preference and raised the contents of neurotransmitters dopamine (DA) and norepinephrine (NE). CGD treatment led to a reduction in lung index and the relative expression of the M gene, along with improvements in pathological changes caused by IAV. Furthermore, the levels of cytokines were decreased after treatment with CGD, and the imbalance in the ratio of Th17/Treg was corrected via a reduction in the number of Th17 cells after treatment.

Conclusions: CGD has a therapeutic effect in mice with depression-like symptoms infected with IAV, increasing the levels of DA and NE, downregulating M gene expression, calming the cytokine storm, and preserving the homeostasis of Th17/Treg cells.

Background: The traditional Sox10^Dom/+ mouse breeding strategy is costly and time-consuming, so this study aims to optimize the breeding method and improve the scientific research efficiency.

Methods: We select the offspring from mating B6C3Fe Sox10^Dom/+ male mice with C57BL/6J female mice, and name the progeny B6C3Fe-g. Further, conduct separate self-breeding for both the B6C3Fe and B6C3Fe-g strains, adhering to the principle of pairing mutants with non-mutants. By comparing the number of offspring, survival rates, and the phenotype of aganglionosis in the colon, a comprehensive evaluation of their breeding capacity and phenotypic stability is conducted.

Results: Sanger sequencing results show that the mutation sites of B6C3Fe and B6C3Fe-g mice are consistent. After fluorescent staining of intestinal nerves, it was found that the heterozygous mice of the two strains had neuronal deletion in the distal colon, and this pathological phenotype was consistent with the pathological features of the diseased colon of Hirschsprung disease (HSCR). However, compared with the B6C3Fe strain, the B6C3Fe-g strain has a higher number of offspring and greater survival rates.

Conclusions: The breeding strategy of the B6C3Fe-g strain ensures genetic and phenotypic stability, while improving reproductive efficiency, and is an ideal scheme for breeding Sox10^Dom/+ mice.

Background: Acute pancreatitis (AP) is a severe inflammation of the pancreas, marked by elevated enzyme levels, cellular inflammation, and necrosis. Recent studies emphasize the critical role of inflammation in AP progression. Tirzepatide, a multi-target agonist of glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) receptors, has demonstrated notable anti-inflammatory and metabolic benefits.

Methods: This study explores the therapeutic potential of Tirzepatide in pancreatitis induced by L-arginine in rats, focusing on enzymatic markers, cytokine profiles, oxidative stress, and histological outcomes. Over 27 days, rats were distributed into Control, Tirzepatide, L-Arginine, and L-Arginine + Tirzepatide groups, with the latter receiving L-Arginine to induce pancreatitis followed by Tirzepatide administration.

Results: L-Arginine significantly elevated serum amylase, lipase, and inflammatory mediators (IL-6, IL-4, and IL-10), alongside oxidative stress markers and histopathological deterioration. Conversely, the L-Arginine + Tirzepatide group exhibited reduced lipase and IL-6 levels, suppressed reactive oxygen species (ROS) generation, and enhanced anti-inflammatory cytokines IL-4 and IL-10. Histopathological analysis revealed reduced necrosis and tissue damage in the L-Arginine + Tirzepatide group compared to the L-Arginine group, indicating Tirzepatide's possible protective effects. Immunofluorescence studies further demonstrated increased p-Akt expression, supporting the role of Tirzepatide in cellular repair and recovery.

Conclusion: These findings highlight Tirzepatide's ability to mitigate pancreatic damage through antioxidant and anti-inflammatory mechanisms, underscoring its potential as a pharmacological agent for acute pancreatitis.

Background: Pulmonary hypertension (PH) is a life-threatening condition that can be triggered by pulmonary thromboembolism (PTE), which causes abrupt increases in pulmonary artery pressure and resistance. Although Doppler echocardiography is a useful screening tool, its ability to accurately reflect rapid hemodynamic changes during acute PTE remains limited. The Flowire catheter allows for real-time assessment of intravascular flow and may offer better insight into these changes.

Aims: The aims were to investigate changes in pulmonary artery hemodynamics measured using a Flowire catheter and to validate the accuracy of Doppler echocardiography in assessing these changes in dogs with acute pulmonary thromboembolism (PTE).

Methods: Hemodynamic and echocardiographic data were obtained from 10 anesthetized female beagles using a Flowire catheter and echocardiography at three preload conditions: baseline, bolus loading, and an acute pulmonary hypertension state induced by a 300-μm dextran microsphere injection.

Results: With increases in pulmonary artery pressure and pulmonary vascular resistance, the proximal and distal pulmonary artery flow peak measured using the Flowire catheter significantly decreased during the acute pulmonary hypertension period. Echocardiography did not accurately capture these hemodynamic changes and tended to overestimate pulmonary artery flow peak in the distal pulmonary artery.

Conclusion: Doppler echocardiography has limitations in accurately reflecting complex hemodynamic changes during acute PTE. In contrast, Flowire catheterization provides additional and precise local hemodynamic information.

Background: Bacterial pneumonia remains a leading cause of morbidity and mortality worldwide despite the widespread availability of antibiotics. Novel pneumonia therapies and biomarkers are urgently needed to improve outcomes and advance personalized therapy. Using an established baboon model of S. pneumoniae pneumonia, we sought to characterize the temporal dynamics of pneumonia host responses to identify novel potential diagnostic and therapeutic molecular targets.

Methods: We performed whole blood transcriptomics, unbiased proteomics, and peripheral cytokine measurements serially in baboons inoculated with S. pneumoniae (n = 23) or saline (n = 10) and modeled the peripheral blood host response using principal components analysis and complex sparse logistic regression. Differentially expressed genes were analyzed for pathway analysis.

Results: Inoculated animals developed characteristic signs and symptoms of pneumonia. A 39-gene signature was derived that classified S. pneumoniae infection with high accuracy (auROC 0.9 and 0.99 at 24 and 48 h post-inoculation, respectively). Similar performance was observed for 48-h biomarker signatures derived from peripheral blood plasma proteomic and cytokine measurements (both auROC >0.9). The gene signature retained strong diagnostic performance (auROC = 0.88) when transformed to human orthologs and applied to patients with acute respiratory illness (n = 34) or healthy controls (n = 20). Pathway analysis at 48 h identified down-regulation of mitophagy and glucocorticoid signaling in peripheral blood.

Conclusions: We report novel peripheral blood gene and protein expression signatures of S. pneumoniae pneumonia that could improve pneumonia diagnosis and found distinct pathways that may be amenable to modulation. Our findings illustrate how non-human primate models of bacterial pneumonia can successfully translate biomarker discoveries to patients.

Over 550 000 people in the U.S. require hemodialysis for management of end stage renal disease (ESRD). When anatomy restricts fistula creation, arteriovenous grafts (AVG) are implanted. AVGs have poor primary patency and high risk of infection, highlighting a need for better alternatives. Previous AVG large animal models were limited by high complication rates and short-term follow-up. This study investigates the safety and durability of an ovine bilateral carotid-jugular AVG model. Eight female sheep underwent bilateral carotid-jugular AVG implantation via a single longitudinal incision overlying the trachea. Ringed PTFE grafts were anastomosed in a “lazy-S” configuration to provide the laxity needed to prevent avulsion and minimize kinking with neck movement. Post-operatively, sheep were evaluated daily to monitor for complications. Duplex ultrasonography of the grafts was performed at regular intervals out to 6 months to evaluate patency. At 6 months, angiography and duplex was performed followed by explant for gross and histologic analysis. Technical success was achieved in 16 of 16 (100%) graft implants. No major complications, including stroke, anastomotic disruption, infection, wound breakdown, or death occurred. Primary patency of control PTFE grafts was 75% at 6 months, paralleling reported rates in humans. Bilateral carotid-jugular AVG implantation in sheep is a safe and durable model for self-controlled long-term evaluation of AVG conduit technology.

In this study, we aimed to develop an in vivo electrophysiological bone-nerve preparation to record the activity of peripheral sensory neurons that innervate the murine tibia. A small nerve that innervates the tibial marrow cavity was identified in isoflurane-anesthetized C57BL/6 mice, and placed over a platinum hook electrode for extracellular recording. Whole-nerve activity was amplified, filtered and sampled at 20 kHz using PowerLab (ADInstruments). A cannula was placed into the marrow cavity to deliver mechanical stimuli (by pressurizing with injection of saline) and/or capsaicin. Optical stimulation was achieved by application of 473 nm blue light (1 Hz, 0.25–0.5 ms, 0–12.5 mW/mm) to the tibial marrow cavity in Wnt1-Cre; loxP-ChR2 mice. Murine bone afferent neurons responded to high threshold noxious mechanical stimulation, coded for the intensity of mechanical stimulation, could be sensitized by capsaicin, and did not suffer stimulus-evoked fatigue at 10-minute interstimulus intervals. Electrical and optical stimulation within the marrow cavity evoked action potentials with conduction velocities in the Aδ and/or C fiber range. These new approaches to recording the activity of bone afferent neurons will allow us to take advantage of transgenic and optogenetic tools to further our understanding of mechanisms that generate and maintain bone pain in the future.