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 investigate the effect of aerobic exercise with acupuncture needles on the learning and memory ability in rats subjected to middle cerebral artery occlusion/reperfusion(MCAO/R)and to explore the underly-ing mechanisms.Method:Male SD rats were randomly divided into four groups:sham,model,aerobic exercise with acupunc-ture needles,and inhibitor groups(n=12 per group).In the sham group,only the left common carotid artery,in-ternal carotid ar tery,and external carotid artery were exposed without inducing ischemia,while the other three groups underwent MCAO/R modeling.To establish an MCAO/R model with reduced NLRP3 expression,rats in the inhibitor group received an intraperitoneal injection of MCC950 60 minutes before modeling.On the first day of post-modeling,the Zea-Longa score and T2 WI were used to screen the MCAO/R rats.From the second day,rats in the aerobic exercise with acupuncture needles and the inhibitor groups were underwent a 14-day intervention.The acupoints were selected for"Shenting"(GV 24)and"Baihui"(GV 20),with sparse waveform at a frequency 2/10Hz and a current intensity of 1-3mA.Simultaneously,the rats performed tread-mill exercise,with speeds of 4m/min(day 1-3),6m/min(day 4-7),and 8m/min(day 8-14)for 30min/day.Afterthe intervention,neurological function was reassessed,and the new object recognition test was used to measure the learning and memory ability,and the pathological changes of the hippocampus were observed by HE staining.The protein expressions of NLRP 3,GSDMD,and Caspase-1 were detected by Western Blot and immunofluorescence,ELISA detected the contents of ILL-1β and IL-18,the expression and localization of IBA 1 and NLRP 3 were detected by double immunofluorescent staining.Result:The hippocampal neurons in the ischemic side of the rats were loosely and disorderly arranged in the model group.Compared to the sham group,the model group showed significantly impaired neurological func-tion and novel object recognition ability(P＜0.01),with significant increased expression of NLRP 3,GSDMD,and Caspase-1(P＜0.01),and elevated ILL-lβ and IL-18 contents(P＜0.01).In addition,IBA 1 and NLRP 3 fluo-rescence expression were enhanced(P＜0.01),and many IBA 1+/NLRP 3+double-positive cells were observed un-der the microscope(P＜0.01).Compared with the model group,the aerobic exercise with acupuncture needles group showed significant improvements in neurological function and the new object recognition index(P＜0.05,P＜0.01),reduced hippocampal pathological injury,decreased expression of NLRP 3,GSDMD,and Caspase-1 d(P＜0.01)and lower IL-lβ and IL-18 levels(P＜0.05);the activation of microglia and the expression of IBA 1,NL-RP 3 and the number of IBA 1+/NLRP 3+double-positive cells decreased(P＜0.01).The inhibitor group demon-strated even greater improvements in neural function and new object recognition performance than the aerobic exercise with acupuncture needles group(P＜0.05,P＜0.01),with further reductions in NLRP 3,GSDMD,Caspase-1 protein expression(P＜0.01)and lower level of IL-1β,IL-18(P＜0.01,P＜0.05);the microglia activation level,the number,arrangement and morphology of nerve cells were closely resembled those in the sham group,and there was minimal NLRP 3 microglia observed under the microscope(P＜0.01).Conclusion:Aerobic exercise with acupuncture needles at"Shenting"(GV 24)and"Baihui"(GV 20)points could improve the learning and memory function in MACO/R rats by inhibiting microglia pyroptosis and activa-tion,thereby reducing neuroinflammation.

OBJECTIVES: Through the investigation of the microhardness and microstructure of permanent tooth enamel at various eruption stages during childhood, this research offers references for the early prevention of childhood dental caries. METHODS: Forty-five premolars extracted due to orthodontic reasons were collected and screened. These premolars were divided into three experimental groups according to the time since eruption: Group A (erupted for 0-1 year), Group B (erupted for 1-3 years), and Group C (erupted for 3-5 years). Additionally, the third molars that were extracted due to impaction and had not erupted were selected as the control group, with 15 teeth in each group. Samples were prepared, and the surface microhardness, microstructure, and elemental composition of the enamel were measured using Vickers microhardness tester, scanning electron microscope, and electron probe, respectively. RESULTS: Compared with that in the control group, the microhardness of enamel in groups A, B, and C increased with prolonged eruption time, the surface porosity structure decreased considerably, the contents of Na and Mg on the surface decreased, and that of F increased (P<0.05). CONCLUSIONS The microhardness and microstructure of enamel in permanent teeth at different stages vary. Permanent teeth are at a substantially higher risk of caries within one year after eruption, and early prevention should be emphasized.
Dental Enamel/ultrastructure* ; Humans ; Hardness ; Dental Caries/prevention & control* ; Microscopy, Electron, Scanning ; Tooth Eruption ; Bicuspid/chemistry* ; Dentition, Permanent ; Child ; Child, Preschool

Objective:To investigate the effect of aerobic exercise with acupuncture needles on the learning and memory ability in rats subjected to middle cerebral artery occlusion/reperfusion(MCAO/R)and to explore the underly-ing mechanisms.Method:Male SD rats were randomly divided into four groups:sham,model,aerobic exercise with acupunc-ture needles,and inhibitor groups(n=12 per group).In the sham group,only the left common carotid artery,in-ternal carotid ar tery,and external carotid artery were exposed without inducing ischemia,while the other three groups underwent MCAO/R modeling.To establish an MCAO/R model with reduced NLRP3 expression,rats in the inhibitor group received an intraperitoneal injection of MCC950 60 minutes before modeling.On the first day of post-modeling,the Zea-Longa score and T2 WI were used to screen the MCAO/R rats.From the second day,rats in the aerobic exercise with acupuncture needles and the inhibitor groups were underwent a 14-day intervention.The acupoints were selected for"Shenting"(GV 24)and"Baihui"(GV 20),with sparse waveform at a frequency 2/10Hz and a current intensity of 1-3mA.Simultaneously,the rats performed tread-mill exercise,with speeds of 4m/min(day 1-3),6m/min(day 4-7),and 8m/min(day 8-14)for 30min/day.Afterthe intervention,neurological function was reassessed,and the new object recognition test was used to measure the learning and memory ability,and the pathological changes of the hippocampus were observed by HE staining.The protein expressions of NLRP 3,GSDMD,and Caspase-1 were detected by Western Blot and immunofluorescence,ELISA detected the contents of ILL-1β and IL-18,the expression and localization of IBA 1 and NLRP 3 were detected by double immunofluorescent staining.Result:The hippocampal neurons in the ischemic side of the rats were loosely and disorderly arranged in the model group.Compared to the sham group,the model group showed significantly impaired neurological func-tion and novel object recognition ability(P＜0.01),with significant increased expression of NLRP 3,GSDMD,and Caspase-1(P＜0.01),and elevated ILL-lβ and IL-18 contents(P＜0.01).In addition,IBA 1 and NLRP 3 fluo-rescence expression were enhanced(P＜0.01),and many IBA 1+/NLRP 3+double-positive cells were observed un-der the microscope(P＜0.01).Compared with the model group,the aerobic exercise with acupuncture needles group showed significant improvements in neurological function and the new object recognition index(P＜0.05,P＜0.01),reduced hippocampal pathological injury,decreased expression of NLRP 3,GSDMD,and Caspase-1 d(P＜0.01)and lower IL-lβ and IL-18 levels(P＜0.05);the activation of microglia and the expression of IBA 1,NL-RP 3 and the number of IBA 1+/NLRP 3+double-positive cells decreased(P＜0.01).The inhibitor group demon-strated even greater improvements in neural function and new object recognition performance than the aerobic exercise with acupuncture needles group(P＜0.05,P＜0.01),with further reductions in NLRP 3,GSDMD,Caspase-1 protein expression(P＜0.01)and lower level of IL-1β,IL-18(P＜0.01,P＜0.05);the microglia activation level,the number,arrangement and morphology of nerve cells were closely resembled those in the sham group,and there was minimal NLRP 3 microglia observed under the microscope(P＜0.01).Conclusion:Aerobic exercise with acupuncture needles at"Shenting"(GV 24)and"Baihui"(GV 20)points could improve the learning and memory function in MACO/R rats by inhibiting microglia pyroptosis and activa-tion,thereby reducing neuroinflammation.

Objective To explore the regulation of Chaihuang Qingyi Huoxue Granule on intestinal microecological changes in rats with severe acute pancreatitis (SAP) and the potential mechanism for its treatment of SAP. Methods Forty-eight rats were randomly divided into sham operation group (SHAM),SAP model group (SAP),and Chaihuang Qingyi Huoxue Granule (CH)group,with 16 rats in each group. Each group was further divided into 12 h and 24 h subgroups. The SAP model was induced by retrograde injection of 5％ sodium taurocholate into the pancreaticobiliary duct through duodenal wall. The SHAM and SAP groups received normal saline by gavage,while the CH group received 1.2 g·kg-1 Chaihuang Qingyi Huoxue Granule solution by gavage every six hours. At 12 h and 24 h after operation,eight rats from each group were sacrificed to collect abdominal aortic blood,pancreatic and ileal tissues for analysis. Ascites,pancreatic and ileal tissues were observed. Serum amylase(AMY) and lipase (LPS) levels were measured biochemically. Pathological changes in pancreatic and ileal tissues were investigated by HE staining. Claudin-1 protein expression in ileal tissue was detected by Western Blot. Changes in the intestinal flora of ileocecal contents were analyzed by 16S rDNA high-throughput sequencing. Results Compared to the SHAM group at the same time points,the SAP group exhibited extensive pancreatic edema and necrosis. Serum AMY and LPS levels,pancreatic and ileal histopathological scores increased,and Claudin-1 protein expression in ileal tissue markedly decreased (all P<0.05). The differences in abundance of microbial community increased,while the evenness of community composition reduced. The microbial richness showed no significant change (P>0.05),but the microbial diversity decreased(P<0.05). Proteobacteria were dominant intestinal bacteria. Relative abundances of Oscillospira,Ruminococcus,Bifidobacterium,and Bacteroides S24-7 decreased,whereas relative abundances of Shigella and Allobaculum increased. The differences in abundance of microbial community reduced,and the evenness of community composition increased. The microbial richness showed no significant change(P>0.05),but the microbial diversity increased (P<0.05). Firmicutes and Bacteroidetes were the dominant intestinal bacteria. Relative abundances of Oscillospira,Ruminococcus,Bifidobacterium,and Bacteroides S24-7 increased,whereas relative abundances of Shigella and Allobaculum decreased. After the intervention of CH,pathological damage in ileal tissue was improved. The expression of Claudin-1 protein in the intestinal mucosal barrier increased compared to the model group(P<0.05). The differences in abundance of microbial community reduced,and the evenness of community composition increased. CH group showed an increase in some beneficial bacteria and decrease in pathogenic bacteria compared to model group. Conclusion Chaihuang Qingyi Huoxue Granule may reduce pancreas injury in rats with SAP,which may be involved in modulating the intestinal microecology and improving intestinal mucosal barrier function.

N6-methyladenosine(m6A)modification is one of the most abundant epitranscriptomic modifications in eukaryotic mRNA,with dynamic and reversible properties.This modification process is coordinated by methyltransferases,demethylases,and related m6A binding proteins,which in turn affect mRNA metabolism and function.Increasing evi-dence has indicated that the m6A RNA modification plays an important role in the occurrence and development of athero-sclerosis(As)and other related diseases.This paper provide a comprehensive review of the relationship between m6A RNA modification and As.The entire manuscript summarizes the m6A RNA modification mechanism and its roles in As-related cells including endothelial cells,macrophages,and smooth muscle cells,and discusses the association of m6A RNA modification with risk factors of As such as high-fat diet,ischemia/hypoxia,oscillatory stress,and hypertension.Finally,this review summarizes researches on drug intervention targeting m6A RNA methylation to mitigate As.These studies pro-vide important references for exploring new targets for early diagnosis and treatment of As.

Lactic acid bacteria(LAB)were isolated and cultivated from the intestinal contents of rabbits by MRS-CaCO3 solid medium.Identification was achieved through morphological observa-tion,Gram staining,physiological and biochemical characterisation,16S rDNA sequence analysis,and ERIC-PCR analysis.Strains displaying typical Lactobacilli characteristics were exanimated for their biological characteristics,resistance properties,adherence capacity in vitro,colonization abili-ty in vivo,and safety profile.In this study,a total of four strains of Lactobacillus reuteri were iso-lated from rabbits,all of which exhibited typical biological characteristics of LAB.These strains demonstrated inhibitory effects on common pathogenic bacteria in the gastrointestinal tract,with the primary inhibitory substance being bacteriocin.Furthermore,they showed sensitivity to chlor-amphenicol,rifampicin,and erythromycin,and displayed a degree of tolerance to gastrointestinal conditions and high temperature.These stains were capable of successful colonization in rabbits with a higher degree of safety.This study lays a foundation for the development of LAB prepara-tions for the prevention and treatment of rabbit intestinal diseases.

Objective To investigate the risk factors for pulmonary embolism (PE)in patients with acute spinal cord injury (ASCI).Methods A case-control design was adopted in this study.Basic information,injury cause,injury site,imaging and laboratory examination results and other relevant data of 840 ASCI patients admitted to 3 affiliated hospitals of Army Medical University from January 2018 to August 2023 were collected.According to occurrence of PE or not,these patients were divided a PE group (n=83) and a non-PE group (n=757).Univariate analysis was performed on 37 risk factors including gender,age,length of hospital stay,length of intensive care unit (ICU)stay,cause of injury,and so on,and then multivariate logistic regression analysis was applied to analyze these selected significant variables.Results Univariate analysis showed that 24 factors such as age,length of hospital stay,lower limb fracture,pulmonary contusion and infection,shock and number of operations were related to the occurrence of PE in ASCI patients (P<0.05).Binary logistic regression analysis indicated that age (P=0.000,OR=1.957,95％CI:1.532～2.499),length of ICU stay (P=0.000,OR=2.561,95％CI:1.728～3.794),number of operations (P=0.015,OR=1.814,95％CI:1.124～2.927),spinal fixation (P=0.004,OR=0.370,95％CI:0.189～0.724),restrictive position (P=0.000,OR=21.269,95％CI:5.276～85.740),platelet count (P=0.000,OR=1.553,95％CI:1.236～1.951),D-dimer level (P=0.000,OR=1.600,95％CI:1.376～1.860)and prothrombin time (P=0.025,OR=2.756,95％CI:1.138～6.670)were independent risk factors for PE in the patients.Conclusion The mortality of the ASCI patients with PE is significantly higher than those without.Age,length of ICU stay,number of operations,spinal fixation,restrictive position,platelet count,D-dimer level and prothrombin time are closely associated with PE in ASCI patients.

Objective:To investigate the efficacy and safety of daratumumab in the treatment of systemic light chain amyloidosis.Methods:The clinical data of 24 patients with systemic light chain amyloidosis who received daratumumab-based regimens in Sichuan Provincial People's Hospital from January 2020 to November 2022 were retrospectively analyzed. The treatment process of patients was summarized and the therapeutic efficacy was evaluated. Kaplan-Meier method was used to make survival analysis and the adverse reactions were analyzed.Results:All 24 patients included 2 cases (8.33%) of Mayo 2004 stageⅠ, 2 cases (8.33%) of Mayo 2004 stage Ⅱ and 20 cases (83.33%) of Mayo 2004 stage Ⅲ. All patients were treated with daratumumab-based regimen, and 17 patients had evaluable efficacy. In the chemotherapy regimens, 15 patients received DVd (daratumumab + bortezomib + dexamethasone) regimen, 7 patients received DVCd (daratumumab + bortezomib + cyclophosphamide + dexamethasone) regimen, 1 patient received DRd (daratumumab + lenalidomide + dexamethasone) regimen, and 1 patient received DTd (daratumumab +thalidomide + dexamethasone) regimen. After 1 course of daratumumab-based regimens in 17 cases with evaluable efficacy, the strict complete remission (sCR) rate was 41.18% (7/17), the overall response rate (ORR) was 88.24% (15/17). Among 17 patients who received daratumumab-based chemotherapy regimen as the first-line treatment, sCR rate of 11 cases with evaluable efficacy was 36.36% (4/11) after 1 course of treatment ORR was 90.90% (10/11). Among 5 relapsed/refractory patients, sCR rate of 4 cases with evaluable efficacy was 50.00% (2/4) after 1 course of treatment; ORR was 75.00% (3/4). Among 24 patients, renal involvement was found in 17 patients at the initial diagnosis. After 1 course of daratumumab-based chemotherapy regimen, ORR of 7 cases with evaluable efficacy was 85.71% (6/7), among which 42.86% (3/7) patients with renal involvement had an assessed renal response of very good partial remission (VGPR) or above. At the initial diagnosis, 19 cases had cardiac involvement; ORR of 14 cases with evaluable efficacy was 85.71% (12/14), among which 42.86% (6/14) patients had cardiac response to VGPR or above. After daratumumab-based chemotherapy regimen, the main adverse reactions were infusion-related adverse reactions, myelosuppression and infection, all of which were tolerated by the patients. The median follow-up time of 24 patients was 7.0 months (0.5- 16.5 months), the median progression-free survival time was 7.0 months (0.5-16.5 months) and the median overall survival time was 7.0 months (0.5-35.0 months).Conclusions:Daratumumab-based chemotherapy regimen has good efficacy and safety in the treatment of systemic light chain amyloidosis.

With the development of neuroimaging technology, cerebral small vessel disease has become a hot research topic in recent years. It has been clearly related to cognitive decline, dementia and gait instability. However, recent studies have found that small cerebral vascular disease with white matter hyperintensities is the main cause of chronic dizziness in the elderly, but the pathogenesis is not completely clear, which may be related to brain neural network disconnection, visual dependence, eye movement disorder caused by abnormal brain tissue structure, oxidative stress regulation disorder, cerebral blood flow self-regulation disorder, and the interaction mechanism between vestibular system and emotional disorder.