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*

Objective:To investigate the patterns of mutation evolution in patients with acute myeloid leukemia (AML) during treatment and the possible clinical significances.Methods:A retrospective case series study was conducted. A total of 103 AML patients who were hospitalized at the Affiliated Yuebei People's Hospital of Medical College of Shantou University from November 2019 to August 2021 and underwent high-throughput next-generation sequencing (NGS) technology to detect the mutations of 128 AML-related genes in bone marrow samples were selected. Based on the NGS results, the somatic gene mutations in samples of patients collected at initial diagnosis (73 cases), complete remission (CR) (30 cases), non-remission (NR) (23 cases), and recurrence (12 cases) were analyzed, and the targeted drugs involved in the gene mutations detected in NR and recurrence samples were summarized.Results:The median age [ M ( Q1, Q3)] of onset for 103 patients was 58 (48, 66) years, including 64 males (61%) and 39 females (39%); 86 cases (83%) were primary AML, and 17 cases (17%) were secondary AML; at the initial diagnosis, 51 cases (50%) had normal karyotypes, 34 cases (33%) had abnormalities, and 18 cases (17.5%) were unknown. Compared with the CR samples, the mutation frequencies of FLT3 [29% (21/73) vs. 3% (1/30)], NPM1 [27% (20/73) vs. 3% (1/30)], NRAS [22% (16/73) vs. 3% (1/30)], and IDH2 [14% (10/73) vs. 0 (0/30)] were all higher in the initial diagnosis samples, and the differences were statistically significant (all P < 0.05); compared with the initial diagnosis sample, the median number of gene mutations in each CR sample was lower [4 (2, 5) vs. 7 (5, 9)], and the difference was statistically significant ( P < 0.001). However, there was no statistically significant difference in the median number of gene mutations in each patient between the initial diagnosis samples and the NR samples, the initial diagnosis samples and the recurrence samples, and the NR samples and the recurrence samples (all P > 0.05). Analysis of 14 patients with NGS data at initial diagnosis and CR showed that the same gene mutations could be detected at initial diagnosis and CR, such as DNAH23 (3 cases), USH2A (3 cases), etc; partial gene mutations were detected at initial diagnosis but were not detected at CR, including NRAS (5 cases), FLT3 (3 cases), ANKRD26 (3 cases), NPM1 (3 cases), ETV6 (3 cases), etc; ARID1B (1 case) and DNMT3A (1 case) were negative for mutations at initial diagnosis but positive upon reaching CR. Analysis of 14 patients with NGS data at initial diagnosis and NR showed that most gene mutations persisted at initial diagnosis and NR, such as DNMT3A (5 cases), NRAS (5 cases), KRAS (3 cases), RUNX1 (3 cases), etc; the mutant genes detected at initial diagnosis but not detected at NR included USH2A (2 cases), PCLO (2 cases), ATM (2 cases), FAT1 (2 cases), etc; partial gene mutations were not detected at initial diagnosis but were detected at NR, such as FAT1 (2 cases), TCF3 (2 cases), etc. Analysis of 5 patients with NGS data at CR and recurrence showed that some gene mutations were detected at both CR and recurrence, such as BCORL1 (1 case), ARID2 (1 case), SETD2 (1 case), VEGFC (1 case), etc; FLT1 (1 case) and GNAS (1 case) gene mutations were detected at CR but not detected at recurrence; at recurrence, some gene mutations that were not detected at CR were also detected, such as ANKRD26 (1 case), WT1 (1 case), etc. Among the 23 NR samples and 12 recurrence samples, the targets of drugs approved by US Food and Drug Administration or in clinical trials were detected in 14 (61%) and 5 (42%) samples respectively, including IDH1, IDH2, FLT3, KIT, KRAS, NRAS, SF3B1, U2AF1, and SRSF2. Conclusions:The number of gene mutations in AML patients during CR is significantly less than that at initial diagnosis, some gene mutations disappear when CR is achieved through treatment, but the majority of gene mutations persist during the treatment period, including NR and recurrence, suggesting that monitoring through NGS technology can help understand the evolution of gene mutations during AML treatment and discover the potential therapeutic targets.

Objective Zinc finger protein 335 (Zfp335) plays a crucial role in the early development of thymic T cells and the differentiation of peripheral T cell subpopulations. The objective of this study is to investigate the role and underlying mechanisms of Zfp335 in the regulation of regulatory T cell (Treg) within tumor immunity. Methods The Zfp335 gene was specifically knocked out in Treg using tamoxifen (Zfp335^fl/fl FOXP3^creERT2), and the MC38 tumor model was established. On the 7th day after tumor inoculation, tumor size was observed and measured. Tumor size was monitored and recorded daily starting from day 7 post-inoculation. On day 12, tumors were harvested, and the proportions of CD4⁺ T cells, CD8⁺ T cells, and Treg were analyzed by flow cytometry. Additionally, the mitochondrial function of effector regulatory T cell (eTreg) was assessed. Results From day 10 post-tumor inoculation, tumor volume in the Zfp335^CKO group was significantly reduced compared to that of the wild-type (WT) group. Furthermore, the infiltration of CD4⁺ and CD8⁺ T cells, along with their respective effector cells, was significantly higher in the Zfp335^CKO group than in the WT group. The proportions of CD4⁺ and CD8⁺ T cells producing interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were also significantly increased in the Zfp335^CKO group compared to that of the WT group. In addition, the percentage of CD8⁺ T cells secreting granzyme B (GzmB) was significantly higher in the Zfp335^CKO group than that in the WT group. In contrast, the proportion of Treg and inducible T cell co-stimulator (ICOS)⁺ Treg in the Zfp335^CKO group was significantly lower than that in the WT group. Finally, the expression level of Mitotracker Deep Red in eTreg from the Zfp335^CKO group was significantly reduced compared to that in the WT group. Conclusion During tumorigenesis, the specific deletion of Zfp335 impairs Treg activation, which is related to decreased mitochondrial function in eTreg. In Zfp335^CKO mice. Tumors exhibit increased infiltration of effector T cells, accompanied by elevated levels of cytotoxic cytokines, ultimately enhancing resistance to tumor progression.
Animals ; T-Lymphocytes, Regulatory/metabolism* ; Mice ; CD8-Positive T-Lymphocytes/immunology* ; Neoplasms/genetics* ; Cell Line, Tumor ; Mice, Inbred C57BL ; Mice, Knockout ; DNA-Binding Proteins/genetics* ; Female

Extracellular vesicles (EVs) are membrane-bound vesicles secreted by cells, including exosomes, microvesicles, and apoptotic bodies, which play critical roles in intercellular communication, material transport, and signal transduction. In recent years, increasing evidence has highlighted the essential function of EVs in early embryo development. By carrying bioactive molecules such as proteins, nucleic acids (e.g., mRNA and miRNA), and lipids, EVs regulate embryonic gene expression, cell proliferation, differentiation, and the microenvironment. Studies have shown that EVs derived from various segments of the female reproductive tract can enhance embryonic developmental potential, improve embryo quality, and facilitate implantation. Additionally, EVs secreted by embryos themselves participate in intercellular communication and play pivotal roles during embryogenesis. This review summarizes recent advances in understanding the functions of EVs in early embryo development, discusses their roles in mediating cell-cell communication and regulating gene expression, and explores their potential applica-tions in reproductive medicine and clinical practice, offering new perspectives for optimizing assisted reproductive technologies.

Autism Spectrum Disorders (ASDs) are reported as a group of neurodevelopmental disorders. The structural changes of brain regions including the hippocampus were widely reported in autistic patients and mouse models with dysfunction of ASD risk genes, but the underlying mechanisms are not fully understood. Here, we report that deletion of Trio, a high-susceptibility gene of ASDs, causes a postnatal dentate gyrus (DG) hypoplasia with a zigzagged suprapyramidal blade, and the Trio-deficient mice display autism-like behaviors. The impaired morphogenesis of DG is mainly caused by disturbing the postnatal distribution of postmitotic granule cells (GCs), which further results in a migration deficit of neural progenitors. Furthermore, we reveal that Trio plays different roles in various excitatory neural cells by spatial transcriptomic sequencing, especially the role of regulating the migration of postmitotic GCs. In summary, our findings provide evidence of cellular mechanisms that Trio is involved in postnatal DG morphogenesis.
Animals ; Dentate Gyrus/metabolism* ; Mice ; Morphogenesis/physiology* ; Neurons/pathology* ; Cell Movement ; Mice, Inbred C57BL ; Autism Spectrum Disorder/pathology* ; Mice, Knockout ; Neural Stem Cells ; Male ; Neurogenesis ; Autistic Disorder/genetics*

To summarise the clinical experience of Professor TONG Xiaolin in treating prediabetes combined with overweight or obesity using the method of relieving depression and reducing turbidity. It is believed that prediabetes belongs to the category of "spleen-heat syndrome" in traditional Chinese medicine， and its core pathogenesis is center fullness with internal heat， while obesity is the initiating factor for exacerbating center fullness and internal heat， therefore， it is of great significance to reduce the risk of diabetes by interrupting the transformation between overweight， obesity and glucose metabolism abnormality. It is proposed that the main pathogenesis of prediabetes combined with overweight or obesity is qi depression and turbidity obstruction in middle jiao， with qi depression as the root and turbidity obstruction as the cause， forming a treatment idea with the method of relieving depression and reducing turbidity as the core. In clinic， Dahuang Huanglian Xiexin Decoction （大黄黄连泻心汤） is used as the basic prescription， with a primary focus on directing the turbid downward， supplemented by regulating qi， which embodies the concept of "promoting movement through descent， then figuring out the root of spleen-heat syndrome. Furthermore， the treatment is flexibly modified based on the patient's deficiency-excess syndrome to ensure individualized therapy.

With the continuous development of refractive surgery, people's focus has gradually shifted from improving vision to improving visual quality, and personalized laser-assisted in situ keratomileusis(LASIK)surgery has gradually become people's preferred choice. Femtosecond laser-assisted keratoplasty provides better advantages for personalized LASIK surgery. This article mainly introduces the commonly used femtosecond laser-assisted personalized LASIK surgery(FS-LASIK)in recent years, such as wavefront-optimized, wavefront-guided, topography-guided, Q value-guided(aspheric cutting), personalized surgery “Wavelight Plus” and personalized surgery when correcting patients with age-related inadequate accommodation. This article focuses on analyzing the advantages and disadvantages of different personalized FS-LASIK, as well as the research progress in recent years, and also focuses on comparing the differences between different personalized surgeries.

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Objective To review the application scope of risk communication based on eHealth technology in the primary prevention of cardiovascular diseases,so as to provide references for future research and application.Methods A systematic search was conducted in PubMed,Cochrane Library,Embase,Web of Science,CINAHL,PsycINFO,CNKI,Wanfang database,and SinoMed.The search time limit was from the establishment of databases to March 1,2024.The included literature was summarized and analyzed.Results A total of 24 articles were included.The forms of eHealth technology applied in risk communication for primary prevention of cardiovascular include online websites,computerized decision support systems,electronic health records,mobile applications,email,telephone,and text messages.The functions include risk assessment,risk notification,personalized advice,risk tracking and reminders.The outcome indicators include risk perception,physiological indicators,lifestyle and behavior,psychological indicators,feasibility evaluation,decision correlation,doctor-patient communication,intention,risk score,and physician drug prescription.Conclusion The use of eHealth technology in risk communication of cardiovascular diseases has potential value in improving patients'risk perception and promoting healthy behaviors.It is necessary to continuously improve the functions of eHealth technology and enhance its precision and intelligence,so as to better meet the needs of medical staff and patients and promote the efficient implementation of primary prevention of cardiovascular diseases.

OBJECTIVE To investigate α1-adrenergic receptors(α1-AR)distribution in mouse tissues and its function on dexmetomidine(DMED)induced toxic effects.METHODS ① Real-time fluorescence quantita-tive PCR was used to detect the relative expression of α1A-AR,α1B-AR,α1D-AR,α2A-AR,α2B-AR and α2C-AR mRNAs in the heart,apical potion of heart,lungs,apical potion of lung,liver,kidneys,abdominal aorta,prefrontal cortex,hippocampus,striatum,brainstem,thalamus,olfactory bulb,and the rest of the brain tissues of the mouse.The relative expression of mRNA were analyzed.② C57BL/6J mice were pretreated with α2 adrenergic receptor antagonist atipamezole ATI(0.005,0.010,0.020,0.025,0.040,0.050 mg·kg-1,im),orα1-adrenoceptor antagonist prazosin(1 mg·kg-1,im)for 15 min,and then DMED(0.20 mg·kg-1,iv)was given to observe the rate of the loss of righting reflex and the immobilization time in mice.③ C57BL/6J mice were treated with DMED(16.38,20.48,25.60,32.00,40.00,and 50.00 mg·kg-1,iv)to observe the lethality of the mice in 24 h.The dose-effect relationship curves of the lethality rate and the half lethal-dose(LD50)were detected.ATI(1,2,4,and 8 mg·kg-1,im)or prazosin(1 mg·kg-1,im)were pretreated 15 min followed by the administration of DMED(25.60 mg·kg-1,iv).The lethality of the mice were recorded for 24 h.HE staining to observe the lung tissue damage in the mice.RESULTS ① The mRNA expression levels of three α1-AR subtype were higher than those of α2-AR subtype.α2A-AR and α2C-AR were highly expressed in the central nervous system.α2B-AR was highly expressed in the brainstem and peripheral tissues.The mRNA expres-sion levels of α1-AR subtypes were higher than those of α2-AR subtypes in heart,apical potion of heart or lung(P＜0.05).② ATI(0.005 to 0.05 mg·kg-1,im)dose dependently antagonized the loss of righting reflex and decreased the immobilization time induced by DMED(0.20 mg·kg-1,iv).In contrast,prazosin(1 mg·kg-1,im)had no effect on the loss of righting reflex induced by DMED(0.20 mg·kg-1,iv).③ The LD50 of DMED in mice was 26.734 mg·kg-1(iv)with a 95％Cl of 23.606-30.000 mg·kg-1.DMED(25.6 mg·kg-1)was selected for subsequent toxicity.ATI(1,2,4,and 8 mg·kg-1,im)did not antagonize the lethality induced by DMED(25.6 mg·kg-1,iv).The high dose of ATI resulted in elevated death rate and accelerated mortality induced by DMED(25.6 mg·kg-1,iv)in the mice.However,prazosin(1 mg·kg-1,im)reduced the lethality of DMED(25.6 mg·kg-1,iv)(P＜0.01).After administration of DMED(25.6 mg·kg-1),the mice lungs showed significant congestion.HE staining of lung tissues revealed obvious vascular hemorrhage,alveolar rupture,and erythrocyte spillage.Prazosin(1 mg·kg-1,im)effectively attenuated the tissue damage in the lungs,but ATI(1 mg·kg-1,im)aggravated the pulmonary hemorrhage.CONCLU-SIONS In cardiopulmonary tissues,the high expression levels of α1 adrenoceptor overactivation,might related with the lethality induced by DMED.