RNA-based gene therapy has been widely used for various diseases, and extensive studies have proved that suitable delivery routes greatly help the development of RNA therapeutics. Identifying a safe and effective delivery system is key to realizing RNA therapeutics' clinical translation. Inhalation is a non-invasive pulmonary delivery modality that can enhance the retention of therapeutic agents in the lungs with negligible toxicity, thereby improving patient compliance. Inhaled RNA therapeutics are increasingly becoming an area of focus for researchers; however, only several clinical trials have explored inhaled delivery of RNA for pulmonary diseases. This review presents an overview of recent advances in inhaled delivery systems for RNA therapeutics, including viral and nonviral systems, highlighting state of the art regarding inhalation in the messenger RNA (mRNA) field. We also summarize the applications of mRNA inhalants in infectious and other lung diseases. Simultaneously, the research progresses on small interfering RNAs (siRNAs), antisense oligonucleotides (ASOs), and different types of RNA are also discussed to provide new strategies for developing RNA inhalation therapy. Finally, we clarify the challenges inhaled RNA-based therapeutics face before their widespread adoption and provide insights to help advance this exciting field to the bedside.

OBJECTIVES: To investigate the effect of orexin-A-mediated regulation of ionotropic glutamate receptors for promoting motor function recovery in rats with spinal cord injury (SCI). METHODS: Thirty-six newborn SD rats (aged 7-14 days) were randomized into 6 groups (n=6), including a normal control group, a sham-operated group, and 4 SCI groups with daily intrathecal injection of saline, DNQX, orexin-A, or orexin-A+DNQX for 3 consecutive days after PCI. Motor function of the rats were evaluated using blood-brain barrier (BBB) score and inclined plane test 1 day before and at 1, 3, and 7 days after SCI. For patch-clamp experiment, spinal cord slices from newborn rats in the control, sham-operated, SCI, and SCI+orexin groups were prepared, and ventral horn neurons were acutely isolated to determine the reversal potential and dynamic indicators of glutamate receptor-mediated currents under glutamate perfusion. RESULTS: At 3 and 7 days after SCI, the orexin-A-treated rats showed significantly higher BBB scores and grip tilt angles than those with other interventions. Compared with those treated with DNQX alone, the rats receiving the combined treatment with orexin and DNQX had significantly higher BBB scores and grip tilt angles on day 7 after PCI. In the patch-clamp experiment, the ventral horn neurons from SCI rat models exhibited obviously higher reversal potential and greater rise slope of glutamate current with shorter decay time than those from sham-operated and orexin-treated rats. CONCLUSIONS Orexin-A promotes motor function recovery in rats after SCI possibly by improving the function of the ionotropic glutamate receptors.
Animals ; Spinal Cord Injuries/drug therapy* ; Rats ; Rats, Sprague-Dawley ; Receptors, Ionotropic Glutamate/metabolism* ; Recovery of Function/drug effects* ; Orexins/pharmacology* ; Male ; Female ; Animals, Newborn ; Neuropeptides/pharmacology* ; Intracellular Signaling Peptides and Proteins/pharmacology*

Compared with conventional transdermal drug delivery systems, dissolving microneedles significantly enhance drug bioavailability by penetrating the stratum corneum barrier and achieving intradermal drug delivery. In order to improve the transdermal bioavailability of dexmedetomidine hydrochloride, in this study, a novel microneedle delivery system was developed for dexmedetomidine hydrochloride based on 3D printing combined with micro-molding. By systematically optimizing the microneedle geometrical parameters, array arrangement, and preparation process parameters, we determined the optimal ratio of drug-carrying matrix as 15% PVP (polyvinyl pyrrolidone) K90. The microneedles exhibited significant drug loading gradients, with mean content of (209.99±27.56) μg/patch, (405.31±30.31) μg/patch, and (621.61±34.43) μg/patch. They showed a regular pyramidal structure under SEM and handheld electron microscopy, and their mechanical strength allowed effective penetration into the stratum corneum. The surface contact angles were all < 90°, indicating excellent hydrophilicity. The microneedles dissolved completely within 10 min after skin insertion, achieving a cumulative release rate of 90% (Higuchi model, r=0.996) during 2 hours of in vitro transdermal permeation. The cytotoxicity test and hemolysis test verified good biocompatibility. Pharmacodynamic evaluation showed that the microneedle group demonstrated pain-relieving effect within 15 min, with the pain threshold at the time point of 60 min being 3 times that in the transdermal cream group. The microneedle system developed in this study not only offers an efficient drug delivery option for patients but also establishes an innovative platform for rapid percutaneous delivery of hydrophilic drugs, demonstrating significant potential in perioperative pain management.
Dexmedetomidine/pharmacokinetics* ; Printing, Three-Dimensional ; Needles ; Drug Delivery Systems/methods* ; Administration, Cutaneous ; Animals ; Microinjections/instrumentation* ; Skin Absorption ; Skin/metabolism*

Platelet transfusion refractoriness (PTR) is one of the common problems in platelet transfusion, significantly impacting patient clinical outcomes and increasing the demand for allogeneic platelet transfusion. Both immune and non-immune factors contribute to PTR, among which the occurrence mechanism of immune platelet transfusion refractoriness (IPTR) involves humoral and cellular immune processes and is influenced by platelet storage, processing, and the patient's disease, therapy and immune status. This review comprehensively discusses the research related to the factors for alloimmune of IPTR, the mechanism of platelet clearance and its influencing factors. Furthermore, it explores feasible prevention and treatment measures such as platelet compatibility transfusion and clinical treatments. The aim is to provide a systematic cognition for a deeper understanding of the pathological process of platelet alloimmunization and clearance, and to provide a theoretical basis for the construction of precise clinical prevention and treatment strategies for IPTR, as well as to explore feasible research directions in this field in the future.

Objective To observe the effect of Jinshui Liujun decoction on airway lesions in mice with COPD and explore its possible mechanism.Methods 48 C57BL/6 mice were randomly divided into blank group,model group,Jinshui Liujun decoction group and NAC group,with 12 in each group.The COPD mouse model was established by intranasal drip of LPS and smoking,and the corresponding drugs were given intragastric administration for 14 days after the model was established.Observe the general condition of the mice,measure the MV,PEF and PIF of the mice with the small animal lung function instrument,semi quantitatively evaluate the inflammation of the lung tissue,the thickness of the alveolar septum and the thickness of the airway wall with HE staining,and observe the airway mucus secretion and goblet cell proliferation with PAS staining.The content of MPO,SA and Urea in BALF was detected by the kit,and the ratio of SA and Urea was calculated.The content of MUC5AC in BALF was detected by ELISA.The levels of ROS,GSH,GSSG and GR in lung tissue were detected with the kit,and the ratio of GSH and GSSG was calculated.The expression level of CFTR mRNA in lung tissue was detected by qRT-PCR.Western blot was used to detect the expression level of CFTR protein in lung tissue.Results Compared with the control group,the growth of mice in the model group was poor.The body weight at each time point during the modeling period decreased(P<0.01),and the indexes of MV,PEF and PIF decreased(P<0.01).The lung tissue pathological score,alveolar septal thickness,airway wall thickness,airway mucus and goblet cell increased(P<0.01).The levels of SA,SA/Urea,MUC5AC and MPO in BALF increased(P<0.01),and the level of Urea decreased(P<0.01),The levels of ROS and GSSG in lung tissue increased(P<0.01),and the levels of GSH,GSH/GSSG,and GR decreased(P<0.01).The expression levels of CFTR mRNA and protein in lung tissue decreased(P<0.01).Compared with the model group,the growth condition of COPD mice improved,the body weight increased at each time point during the modeling period(P<0.05,P<0.01),the indexes of MV,PEF and PIF improved significantly(P<0.01),the pathological score of lung tissue,the thickness of alveolar septa,the thickness of airway wall,airway mucus and goblet cell decreased(P<0.01,P<0.05),and the levels of SA,SA/Urea,MUC5AC and MPO in BALF decreased(P<0.01),And an increase in Urea levels(P<0.01),a decrease in ROS and GSSG levels in lung tissue(P<0.01),and an increase in GSH,GSH/GSSG,and GR levels(P<0.01).The expression levels of CFTR mRNA and protein in lung tissue increased(P<0.01).Conclusion Jinshui Liujun decoction can correct CFTR acquired defects through antioxidant effects to improve airway lesions in COPD.

Objective:To construct and prepare recombinant virus strains chimeric with human metapneumovirus (HMPV) antigenic epitopes.Methods:Recombinant influenza virus vectors which chimeric with different HMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. The recombinant influenza virus strain used the internal genes of A/PR/8/34 (PB1, PB2, PA, NP, NS, M, HA, and NA) as a backbone, with concomitant genetic modifications to insert the B-cell epitopes of HMPV into the HA gene, and the CTL+ Th cell epitopes of HMPV into the NA gene. Preparation of recombinant influenza virus strains using reverse genetics in a " 7+ 1" model. The recombinant virus strains were evaluated by measuring hemagglutinin (HA) titers, half tissue culture infectious dose (TCID 50) and growth curves. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric HMPV epitopes. Results:The epitopes of HMPV were inserted into the influenza virus genome and two recombinant influenza virus strains were rescued successfully, named as FLU/HMPV/B and FLU/HMPV/CTL+ Th. HA titers of the recombinant strains were both 2 7, their TCID 50 were 10 5.2/ml and 10 5.0/ml, respectively. After cultured for three passages in chick embryo, these two recombinant strains could proliferate steadily. Whole genome sequencing verified that the FLU/HMPV/B carried the B-cell epitopes of HMPV, the FLU/HMPV/CTL+ Th carried the CTL and Th cell epitopes of HMPV. Growth curve tests also verified that the recombinant strains could proliferate steadily in eggs. Conclusions:Two recombinant influenza virus vector strains carrying the B cell, CTL and Th epitopes of HMPV were rescued successfully. The result of the recombinant virus strains in terms of growth characteristics as well as genetic stability indicate that they meet the requirements for proceeding to the next step of animal experiments. The immunogenicity and immunoprotective effect will be further evaluated by mouse experiments. Ultimately new ideas for the realization of " one vaccine for two uses" or " one vaccine formultiple uses", as well as a new strategy for the development of HMPV vaccine will be proposed.

Objective:To identify the factors associated with the care needs of the older adults aged 65-105 by age groups,and to compare these factors across different age groups.Methods:A total of 12 244 older adults from the Chinese longitudinal healthy longevity survey(CLHLS)conducted in 2018 were included in the analyses.The participants were categorized into three age groups:young-old(aged 65-79),middle-old(aged 80-89),and oldest-old(aged 90-105).The level of disability was measured by the disability index(DI)in four dimensions,reflecting their care needs.Potential factors associated with care needs were selected based on the health ecological model(HEM),including per-spectives of personal characteristics,behavioral characteristics,interpersonal network,living and working conditions,and policy environment.Multifactor analysis was performed using multinomial Logistic regres-sion.Results:Among China's 12 244 older adults,43.4％had medium or high care needs.Factors for higher care needs of older adults included higher age,higher number of chronic diseases,no exercise habit,excessive sleep duration(≥9 h/d),depressive tendency,living with children or spouse,and un-educated(all P＜0.05).In addition,the young-old group who were past smokers(OR=2.009,95％CI:1.019-3.959),were past drinkers(OR=2.213,95％CI:1.141-4.291),and reported self-per-ceived poverty(OR=2.051,95％CI:1.189-3.540),had higher level of care needs.The middle-old group who were female(OR=1.373,95％CI:1.038-1.817),never drank alcohol(OR=1.551,95％CI:1.059-2.269),and were lack of medical insurance(OR=1.598,95％CI:1.053-2.426),and had higher level of care needs.The oldest-old group who were female(medium care needs vs.low care needs:OR=1.412,95％CI:1.062-1.878;high care needs vs.low care needs:OR=1.506,95％CI:1.137-1.993),reported self-perceived poverty(OR=2.064,95％CI:1.282-3.323),and were lack of medical insurance(OR=1.621,95％CI:1.148-2.291),and had higher level of care needs.Conclusion:The identical factors associated with care needs across different age groups include age,chronic disease,exercise,sleep,depression,living arrangement,and education.Smoking,alcohol consumption,and economic status are specific factors among the young-old group of the older adults,while gender and medical insurance are specific factors among the middle-old and the oldest-old group of the older adults.We recommend conducting prospective cohort studies and intervention studies among specific age groups on the above factors to provide reliable evidence for policy formulation.

Objective:To analyze the potential biomarkers of kidney yang deficiency, kidney yin deficiency, and stasis and water stagnation in diabetic nephropathies (DN) using hydrogen nuclear magnetic resonance ( 1H-NMR)-based technology; To provide a test basis for the TCM diagnosis and typing of DN from the molecular biology point of view. Methods:This is a case-control study. Totally 90 patients with DN Ⅲ-Ⅳ in the second ward of Nephrology Department of Shanxi Integrated Traditional and Western Medicine Hospitalwere selected from November 2022 to May 2023, and were divided into the kidney yang deficiency group, the kidney yin deficiency group, and the stasis and water stagnation group based on TCM syndrome types, with 30 cases in each group; 30 healthy medical checkups were selected as the healthy control group. Urine samples were taken from each group for case-control study, and the samples were examined by 1H-NMR technology. The data were preprocessed to obtain the relevant data matrix and qualitative identification of metabolites, and multivariate statistical analyses were performed to clarify the differences between the samples and screen the metabolites that contributed to the grouping, and the key pathways were analyzed. Results:Through metabolomics technology combined with multivariate statistical analysis, a total of 12 potential biomarkers in the urine of patients with DN kidney yang deficiency were screened out, involving 6 major metabolic pathways; 15 potential biomarkers in the urine of patients with kidney yin deficiency were screened out, involving 6 major metabolic pathways; and 7 potential biomarkers in the urine of patients with stasis and water stagnation were screened out, involving 5 major metabolic pathways. Among them, glyoxylate and dicarboxylic acid metabolism, and glycine, serine and threonine metabolism were the metabolic pathways commonly involved in the three groups of syndrome types.Conclusion:There are differences among patients with kidney yang deficiency, kidney yin deficiency, and stasis and water stagnation in DN, and the discovery of potential biomarkers can provide an objective basis and reference value for TCM diagnosis of DN.