Local anesthetics (LAs), such as articaine (AT), exhibit limited efficacy in inflammatory environments, which constitutes a significant limitation in their clinical application within oral medicine. In our prior research, we developed AT-17, which demonstrated effective properties in chronic inflammatory conditions and appears to function as a novel oral LA that could address this challenge. In the present study, we further elucidated the beneficial effects of AT-17 in acute inflammation, particularly in oral acute inflammation, where mitochondrial-related apoptosis played a crucial role. Our findings indicated that AT-17 effectively inhibited lipopolysaccharide (LPS)-induced nerve cell apoptosis by ameliorating mitochondrial dysfunction in vitro. This process involved the inhibition of mitochondrial reactive oxygen species (mtROS) production and the subsequent activation of the NRF2 pathway. Most notably, improvements in mitochondria-related apoptosis were key contributors to AT-17's inhibition of voltage-gated sodium channels. Additionally, AT-17 was shown to reduce mtROS production in nerve cells through the Na⁺/NCLX/ETC signaling axis. In conclusion, we have developed a novel local anesthetic that exhibits pronounced anesthetic functionality under inflammatory conditions by enhancing mitochondria-related apoptosis. This advancement holds considerable promise for future drug development and deepening our understanding of the underlying mechanisms of action.

Hypoxic-ischemic (HI) brain damage poses a high risk of death or lifelong disability, yet effective treatments remain elusive. Here, we demonstrated that miR-100-5p levels in the lesioned cortex increased after HI insult in neonatal mice. Knockdown of miR-100-5p expression in the brain attenuated brain injury and promoted functional recovery, through inhibiting the cleaved-caspase-3 level, microglia activation, and the release of proinflammation cytokines following HI injury. Engineered extracellular vesicles (EVs) containing neuron-targeting rabies virus glycoprotein (RVG) and miR-100-5p antagonists (RVG-EVs-Antagomir) selectively targeted brain lesions and reduced miR-100-5p levels after intranasal delivery. Both pre- and post-HI administration showed therapeutic benefits. Mechanistically, we identified protein phosphatase 3 catalytic subunit alpha (Ppp3ca) as a novel candidate target gene of miR-100-5p, inhibiting c-Fos expression and neuronal apoptosis following HI insult. In conclusion, our non-invasive method using engineered EVs to deliver miR-100-5p antagomirs to the brain significantly improves functional recovery after HI injury by targeting Ppp3ca to suppress neuronal apoptosis.
Animals ; MicroRNAs/metabolism* ; Extracellular Vesicles/metabolism* ; Mice ; Recovery of Function/physiology* ; Hypoxia-Ischemia, Brain/therapy* ; Mice, Inbred C57BL ; Antagomirs/administration & dosage* ; Male ; Animals, Newborn ; Apoptosis/drug effects* ; Brain Injuries/metabolism* ; Glycoproteins ; Peptide Fragments ; Viral Proteins

Objective To prepare glucose transporter 1(Glut1)-targeted doxorubicin(DOX)-loaded liposomes(doxorubicin/polyphyllin H-liposomes,DOX/ppH-LPs)using polyphyllin H(ppH)instead of cholesterol as the liposomal membrane material,and to investigate their in vitro synergistic anti-tumor activity against non-small cell lung cancer(NSCLC).Methods DOX/ppH-LPs were prepared using thin-film hydration,and the formulation was optimized by single-factor investigation.The optimized DOX/ppH-LPs were characterized for morphology,particle size,polydispersity index(PDI),and zeta potential with transmission electron microscopy(TEM)and dynamic light scattering(DLS).Drug loading DL%was determined by high-performance liquid chromatography(HPLC).The storage stability was evaluated by observing in PBS at 4℃for 7 d,and the serum stability was observed in DMEM containing 10%fetal bovine serum(FBS)at 37℃for 48 h.In vitro drug release was studied in PBS at pH 7.4 and pH 5.0 values,respectively.Human NSCLC A549 cells were subjected as the model,MTT assay was performed to detect the proliferation inhibition by DOX/ppH-LPs at different concentrations(0.5,5.0,15.0 μg/mL)and the control group(ppH+DOX/LPs,a physical mixture of free ppH and DOX-loaded liposomes).Fluorescence microscopy was used to observe cellular uptake of DOX/ppH-LPs and DOX/LPs(containing 5 μg/mL DOX)at 15 min and 2 h.Live/dead cell staining was applied to assess apoptosis/necrosis induced by formulations(15 μg/mL DOX)after 48 h incubation.Transwell assay was conducted to evaluate inhibitory effect on cell migration and invasion,and the targeting property and in vitro synergistic anti-NSCLC activity of DOX/ppH-LPs were then comprehensively evaluated.Results The optimal formulation of DOX/ppH-LPs was determined as hydration temperature at 50℃,6 mg DOX,2 mg ppH,and 24 mg lecithin.The prepared DOX/ppH-LPs were in spherical shape,uniform distribution,and at an average particle size of 145.13±22.14 nm,a PDI of 0.15±0.05,a zeta potential of-23.92±1.73 mV,and a DL of 10.13±0.71%for DOX and(1.22±0.21)%for ppH.DOX/ppH-LPs maintained stable particle size,PDI,and exhibited significantly unchanged zeta potential after storage in PBS at 4℃for 7 d or incubation in DMEM containing 10%FBS at 37℃for 48 h,demonstrating excellent physical and serum stability.Both liposomes showed slow release at pH 7.4 value,while drug release was significantly accelerated at pH 5.0 value(P<0.05),indicating pH-sensitive release characteristics.MTT assay revealed that DOX/ppH-LPs exerted significantly stronger cytotoxicity against A549 cells than the ppH+DOX/LPs control group(P<0.05).Compared with ppH+DOX/LPs,DOX/ppH-LPs showed remarkably enhanced cellular uptake in A549 cells(P<0.05),with more DOX localized in the nucleus.Live/dead cell staining showed that at the same DOX concentration(15 μg/mL),the proportion of apoptotic/necrotic cells induced by DOX/ppH-LPs was significantly higher than that of the DOX/LPs control group.Transwell assay demonstrated that there were significantly less cells migrating and invading through the membrane in the DOX/ppH-LPs group than the ppH+DOX/LPs group.Conclusion Glut1-targeted doxorubicin-loaded liposomes(DOX/ppH-LPs)constructed by substituting cholesterol with ppH can target NSCLC cells,significantly enhance the in vitro synergistic anti-NSCLC activity of DOX and ppH.

Apurinic/apyrimidinic endonuclease 1(APE 1)is a multifunctional protein that plays important roles in DNA repair and regulation of gene expression.Because APE 1 is overexpressed in various cancers,it can serve as a cancer biomarker for aiding clinical diagnosis,guiding therapy,and monitoring prognosis.On this basis,a label-free fluorescent probe was designed based on the primer exchange reaction(PER)strategy for highly sensitive detection of APE 1 activity.In the absence of APE 1,the structure of catalytic hairpin(HP)was stable and could not form G-quadruplex.Therefore,the background fluorescence of this sensing system was very low due to the dissociation of thioflavin T(ThT).In the presence of APE 1,the apurinic/apyrimidinic(AP)site of HP was cleaved by APE 1 and a short nucleic acid fragment that acted as a primer to initiate PER was generated.After PER reaction,a large number of G-quadruplex were produced,which could specifically bind with ThT and resulted in significant increase of fluorescence signal.The combination of low background design of HP and PER amplification made this biosensor had high sensitivity with a detection limit(3σ)of 0.0008 U/mL.Furthermore,the primer sequence was directly generated by the cleavage of APE 1 without additional addition,which not only increased the specificity of the reaction,but also simplified the experiment procedure.Moreover,the use of label-free fluorescence signal reduced the cost of the experiment,and realized rapid detection of APE 1.Finally,this sensor was used to detect APE 1 in human serum samples with spiked recoveries of 91%-104%,proving great potential in study of biological enzyme.

BACKGROUND:Reconstruction of the medial patellofemoral ligament is the primary and basic treatment for patellofemoral dislocation.Generally,autologous or allogeneic tendons are used to fix the patellofemoral podogram area and the femoral insertion,respectively.There are various fixation methods.Patellar lateral insertion fixation methods are relatively diverse,mainly traditional transosseous fixation and recent anchoring methods,including single tunnel,double-tunnel transosseous fixation,and two-wire anchor fixation.However,which fixation method is more effective in reducing patellofemoral joint stress and is more biomechanical has not been determined. OBJECTIVE:To observe the mechanical effects of patellofemoral joint and medial patellofemoral ligament reconstruction with transosseous and wire anchor fixation. METHODS:A three-dimensional finite element model of the knee joint was constructed.The medial patellofemoral ligament was reconstructed by a single tunnel through the bony canal or two wire anchors at the medial edge of the patella.The femoral side was fixed by extrusion nails to the medial epicondyle of the femur and the midpoint of the adductor tubercle.We observed the effects of the two fixation methods on patellofemoral joint and medial patellofemoral ligament loading during knee flexion at 0°,30°,60°,90°,and 120°. RESULTS AND CONCLUSION:(1)The stress on the patellofemoral joint was large when the knee was at flexion of 0-60°,peaked at 30°,and gradually decreased at 90° and 120°.The two fixation methods had little difference in the stress on the patellofemoral joint.(2)The stress on the medial patellofemoral ligament peaked at 30° and decreased significantly at 60°,and the load on the medial patellofemoral ligament was significantly greater at all angles with anchor fixation than with bony canal fixation.(3)The results showed that there was no significant difference in patellofemoral joint loading between the two fixation methods,but the stress on the medial patellofemoral ligament in anchoring was significantly greater than that in transosseous fixation.

Field epidemiological investigation and treatment of a human skin anthrax outbreak were performed,including tracing of the infection source,case searching,management of close contacts,sampling and testing,and on-site elimination.Additional training and public outreach,and anthrax prevention and control measures are recommended.This case involved a sporadic outbreak of human skin anthrax,with sick cattle as the source of infection.On July 20,our patient slaughtered a cow at his home without taking any protective measures.Three days earlier,he had been stabbed at the second joint of the middle finger of his right hand,and he became infected with anthrax bacilli through the wound.Fever appeared on the day after expo-sure,and on the fourth day(July 24),a black scab mass of approximately(3X3)cm was visible at the wound site,without rupture or discharge of pus.He was hospitalized with fever at the 164th Regiment Hospital,Tacheng People's Hospital,Tacheng District People's Hospital,and Tacheng Jingcheng Hospital.His stay at the Tacheng District People's Hospital was from the 24th to 25th.He had seven close contacts,none of whom were infected,and no common exposures were identified.On the 27th,the PCR results for smear samples around the black scab were positive,the serum antibody results were positive,and the fecal samples were negative.The PCR test results for five environmental specimens were negative,and the isolation and culture results for bone surface specimens from dead cattle were positive.Strengthened efforts are necessary in training and public education regarding zoonotic disease prevention and control;establishment of local corps integration and joint prevention and control;and detection and mitigation of epi-demics in early stages.

Objective To investigate the characteristics of oral and laryngeal diadochokinesis abilities and the developmental patterns of coordinated vocal fold movements in preschool children aged 3～6 years.Methods Sixty-one normal preschool children aged 3～6 years were selected to measure their maximum phonation time(MPT),as well as two types of acoustic parameters of oral and laryngeal diadochokinesis ability movements,including the as-sessment of one trisyllabic/pataka/diadochokinesis ability rate and four monosyllabic/?a/,/ha/,/?A/,/hA/la-ryngeal diadochokinesis ability rates.Results Diadochokinesis ability(DDK)and laryngeal diadochokinesis ability(LDDK)rates increased with age in preschool children aged 3～6 years,showing highly significant differences(P＜0.01).The growth rate of children's LDDK rates at age 4～5 years was the maximum within the 3～6 year age range.Children's MPT was highly significantly and positively correlated with DDK and LDDK(P＜0.01).The vo-cal fold adduction capacity in laryngeal diadochokinesis ability movements in children of all age groups was greater than vocal fold adduction ability/?a/LDDK＞/ha/LDDK(P＜0.01),/?A/LDDK＞/h∧/LDDK(P＜0.05).Con-clusion Age,MPT,and vocal fold adduction and abduction status had a significant effect on the oral and laryngeal diadochokinesis ability of preschool children aged 3～6 years old.The ages of 4～5 is a critical period for the devel-opment of children's oral and laryngeal diadochokinesis ability.

Acute skin failure is a common condition of skin damage in critically ill patients, which seriously affects the prognosis of patients. This article summarizes the evaluation indicators, techniques, and comparison of evaluation indicators and techniques for acute skin failure, in order to provide references for the development of acute skin failure evaluation tools and the formulation of nursing measures.

Objective To compare the biomechanical stability of three cross-bridge headless compression screws and lock-ing plates in the fixation of Mason type Ⅲ radial head fractures by finite element method.Methods Using reverse modeling technology,the radial CT data and internal fixation data of a healthy 25-year-old male were imported into the relevant software.Three-dimensional finite element model of 3 cross-bridge headless compression screws and locking plates for Mason Ⅲ radial head fractures were established,and the radial head was loaded with 100 N axial loading.The maximum displacement,maxi-mum Von Mises stress and stress distribution of the two groups were compared.Results The maximum displacements of the three cross-bridge screws group and locking plate group were 0.069 mm and 0.087 mm respectively,and the Von Mises stress peaks were 18.59 MPa and 31.85 MPa respectively.The stress distribution of the three screws group was more uniform.Con-clusion Both internal fixation methods can provide good fixation effect.CoMPared with the locking plate fixation method,the 3 cross-bridge headless compression screws fixation is more stable and the stress distribution is more uniform.

Pulmonary hypertension(PH)is the pulmonary vascular remodeling caused by endothe-lial dysfunction and other factors,which leads to the increase of pulmonary artery pressure and finally right heart failure and patient death.Medications can significantly improve the symptoms of patients with pulmonary hypertension,but drugs can't cure vascular remodeling and right heart failure once for all,so it is urgent to find new treatments.Endothelial cells can obtain mesenchymal phenotypes after hypoxia stimulation,that is,endothelial-to-mesenchymal transition(EndMT).Increasing studies have shown that EndMT plays an important role in pulmonary vascular remodeling of PH.In this review,three signaling pathways related to induction of endothelial-to-mesenchymal transition under hypoxic conditions and research progress in related drugs are elaborated:① Transforming growth factor-β/bone morphogenetic protein(TGF-β/BMP)signaling pathway.The TGF-β/BMP pathway is mediated by Smad,and hypoxia regulates the expressions of EndMT-related genes by inhibiting or inducing the phosphorylation level of Smad in the downstream medium.② Notch signaling pathway.Hypoxia can enhance Jagged/Notch signaling pathway and promote the EndMT process.③Wnt/β-catenin signaling pathway.Hypoxia increases the expression of β-catenin,activates the Wnt signaling pathway,and regulates the expressions of EndMT regulatory genes.The significance of hypoxia-induced EndMT as an impor-tant pathogenic factor in PH is clarified in order to provide new ideas for the improvement of pulmonary vascular remodeling and recommend new effective strategies for the prevention and treatment of PH.