ObjectiveTo investigate the effects of modified Buyang Huanwu Tang on cerebral ischemia-reperfusion injury （CI/RI） in mice via the PTEN-induced putative kinase 1/E3 ubiquitin ligase （PINK1/Parkin） signaling pathway-mediated mitophagy， and to explore the underlying mechanism by which modified Buyang Huanwu Tang improves CI/RI. MethodsSeventy-two male C57BL/6J mice were randomly divided into six groups （n = 12 per group）： Sham-operated group， middle cerebral artery occlusion/reperfusion （MCAO/R） model group， low-， medium-， and high-dose modified Buyang Huanwu Tang groups （8.84， 17.68， 35.36 g·kg^-1·d^-1）， and an aspirin group （13.00 mg·kg^-1·d^-1）. Neurological deficit scores were assessed using the Zea-Longa method. Cerebral infarct volume ratio was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Histopathological changes and neuronal injury in brain tissues were observed using hematoxylin-eosin （HE） staining and Nissl staining. Apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） assay. Mitochondrial ultrastructure in brain tissue was observed by transmission electron microscopy （TEM）. Serum levels of superoxide dismutase （SOD） and malondialdehyde （MDA） were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of PINK1， Parkin， microtubule-associated protein 1 light chain 3B （LC3B， LC3Ⅱ/Ⅰ）， and p62 in brain tissues were detected by real-time quantitative reverse transcription PCR （Real-time PCR） and Western blot， respectively. ResultsCompared with the sham-operated group， the MCAO/R model group showed significantly increased neurological deficit scores and cerebral infarct volume ratios （P<0.01）. Severe cortical injury on the infarct side was observed， characterized by decreased neuronal density， cytoplasmic vacuolation， nuclear pyknosis， a marked reduction in Nissl bodies， dissolution of Nissl bodies in the cytoplasm of some pyramidal neurons， and blurred cellular boundaries. The number of TUNEL-positive cells increased significantly （P<0.01）. Mitochondria exhibited cristae membrane rupture and matrix vacuolation， with rupture of the outer mitochondrial membrane and formation of autophagosomes， the number of which increased significantly. Serum SOD activity decreased significantly （P<0.01）， while MDA content increased significantly （P<0.01）. In infarcted brain tissues of model mice， the relative mRNA expression and protein levels of PINK1， Parkin and LC3B were significantly increased （P<0.05， P<0.01）， whereas p62 mRNA and protein expression were significantly decreased （P<0.05， P<0.01）， showing statistical significance. Compared with the model group， all treatment groups showed significantly decreased neurological deficit scores and cerebral infarct volume ratios （P<0.01）. Neuronal density increased significantly， cytoplasmic vacuolation was alleviated， nuclear morphology tended to be more regular and clearer， Nissl body density increased significantly with reduced dissolution and improved contour clarity. The mitochondrial cristae structure was partially restored， with some mitochondria showing autophagosome encapsulation， and the degree of mitochondrial damage was alleviated. Serum SOD activity increased significantly （P<0.01）， while MDA content decreased significantly. The mRNA and protein expression levels of PINK1， Parkin， and LC3Ⅱ/Ⅰ were significantly increased （P<0.05， P<0.01）， while p62 mRNA and protein expression in the low- and medium-dose modified Buyang Huanwu Tang groups were significantly decreased （P<0.05， P<0.01）， showing statistical significance. ConclusionModified Buyang Huanwu Tang can upregulate the protein expression levels of PINK1， Parkin， and LC3Ⅱ/Ⅰ and downregulate p62 protein expression， suggesting that it may improve CI/RI by regulating the expression of proteins related to the PINK1/Parkin signaling pathway. Regulation of the mitophagy pathway may be one of the mechanisms by which modified Buyang Huanwu Tang alleviates CI/RI in mice.

Objective: To analyze the association of mother-child interaction and screen exposure of preschool children with psychological and behavioural problems, so as to provide guidance for promoting the psychological development of preschool children. Methods: From November to December 2024, a convenience cluster sampling method was used to survey 2 977 mothers of preschool children in Daguan and Yingjiang districts of Anqing City. The Chinese Parent-Child Interaction Scale (CPCIS) was applied to evaluate the quality of mother-child interaction, and the Conners Parent Symptom Questionnaire (PSQ) was used to assess the psychological and behavioral problems of preschool children. Binary Logistic regression was applied to analyze the association of mother-child interaction, screen exposure and their combined effect on psychological and behavioral problems among preschool children. Results: The detection rate of psychological and behavioral problems among preschool children was 13.9%. Binary Logistic regression results showed that low scores of mother-child interaction ( OR=2.31, 95%CI =1.72-3.11) and high screen exposure ( OR= 1.52 , 95%CI =1.23-1.88) were higher risks for psychological and behavioral problems in preschool children; the results of the combined effect showed that preschool children in low scores of mother-child interaction and low screen exposure group ( OR=2.18, 95%CI =1.46-3.28), low scores of mother-child interaction and high screen exposure group ( OR=3.13, 95%CI =2.10- 4.65 ) had significantly higher risks of abnormal detection in psychological and behavioral problems, compared to those in the high scores of mother-child interaction and low screen exposure group respectively (all P <0.05). Conclusions Both screen exposure and mother- child interaction are associated with psychological and behavioral problems in preschool children. High quality mother-child interaction can relieve the adverse effects of screen exposure on preschool children s psychological and behavioral development.

OBJECTIVE: To compare the protective effects of electroacupuncture (EA) and transcutaneous electrical acupoint stimulation (TEAS) during pregnancy on maternal immune activation (MIA)-induced adverse pregnancy outcomes, fetal developmental defects, and neuropsychological behavior abnormalities in offspring mice. METHODS: Eighty pregnant C57BL/6 mice were randomly divided into 5 groups: control, model, EA, TEAS, and sham-stimulation groups, 16 mice in each group. MIA models were replicated on the day 12.5 of pregnancy via tail intravenous injection with polyinosinic-polycytidylic acid. On the second day of modeling success, in the EA and TEAS groups, the interventions were delivered at bilateral "Zusanli" (ST36), with a frequency of 2 Hz, a current of 0.5 mA, and for 20 min each day in the pregnant mice; and the interventions lasted 6 days. Body mass and fertility indexes of pregnant mice, and the development indexes of offspring mice were recorded. Liquid phase suspension chip technology was used to detect the levels of cytokines and chemotactic factors in the serum of pregnant mice and and fetal brain of offspring mice. Flow cytometry was adopted to detect the proportion of the subgroups and subtypes of spleen T lymphocytes and macrophages in pregnant mice. Using the open field test, prepulse inhibition (PPI) test and Morris water maze, the spatial learning and memory were assessed in offspring mice. Immunofluorescence staining was used to detect microglial count in the medial prefrontal cortex (mPFC) in offspring mice. RESULTS: Compared with the control group, the model group showed a reduced body mass of pregnancy mice (P<0.01), smaller litter size and fewer live births (P<0.01, P<0.05), the increase in dead birth and the decrease in offspring survival rate (P<0.05, P<0.01). When compared with model group, in the EA group and the TEAS group, the body mass of pregnancy mice rose (P<0.05), litter size and live births increased (P<0.05, P<0.01), the dead birth was reduced and the offspring survival rate higher (P<0.05). In comparison with the control group, the model group showed the increase in the levels of monocyte chemotactic protein-1 (MCP-1), interleukin-6 (IL-6), γ-interferon (IFN-γ) in the serum of pregnant mice, and spleen M1 macrophage proportion (P<0.01, P<0.05), and the decrease in spleen M2 macrophages of pregnant mice (P<0.01); and the increase in MCP-1 and IL-6 in fetal brain of offspring mice (P<0.05). Compared with the model group, the EA group and the TEAS group showed the decrease in MCP-1, IL-6 and IFN-γ, and spleen M1 macrophage proportion (P<0.01, P<0.05), and the increase in spleen M2 macrophages of pregnant mice (P<0.01, P<0.05) ; and the decrease in MCP-1 and IL-6 in fetal brain of offspring mice (P<0.05). Compared with the control group, in the model group, the total movement distance, escape incubation were extended (P<0.05, P<0.01), the frequency of entering the central area and crossing the platform decreased, and the activity duration in central area was shortened (P<0.05, P<0.01), the average speed rose (P<0.05), PPI%, the percentage of target quadrant swimming time in the total time and that of target quadrant swimming distance in the total distance were reduced (P<0.05, P<0.01) in offspring mice. When compared with the model group, in the EA group and TEAS group, the total movement distance and escape incubation were shortened, the average speed was reduced (P<0.05), PPI% and the frequency of crossing the platform increased (P<0.05, P<0.01); the percentage of target quadrant swimming time in the total time and that of target quadrant swimming distance in the total distance rose (P<0.05, P<0.01) in the offspring mice. In the EA group, the frequency of entering the central area and the activity duration in central area were higher (P<0.05, P<0.01); and in the the TEAS group, the activity duration in central area were longer (P<0.05). When compared with the control group, in the model group, microglial count in mPFC was elevated in offspring mice (P<0.05). In comparison with the model group, the EA group and the TEAS group showed the decrease of microglial count in mPFC (P<0.05). CONCLUSION EA and TEAS at "Zusanli" (ST36) during pregnancy effectively improve in the pregnancy outcomes and fetal brain developmental abnormalities induced by infection, and attenuate neurodevelopmental defects and mental disorders of offspring mice through inhibiting inflammatory activation of microglia in mPFC.
Animals ; Female ; Pregnancy ; Electroacupuncture ; Acupuncture Points ; Mice ; Mice, Inbred C57BL ; Humans ; Male

The ambiguity of etiology makes temporomandibular joint osteoarthritis (TMJOA) "difficult-to-treat". Emerging evidence underscores the therapeutic promise of exosomes in osteoarthritis management. Nonetheless, challenges such as low yields and insignificant efficacy of current exosome therapies necessitate significant advances. Addressing lower strontium (Sr) levels in arthritic synovial microenvironment, we studied the effect of Sr element on exosomes and miRNA selectively loading in synovial mesenchymal stem cells (SMSCs). Here, we developed an optimized system that boosts the yield of SMSC-derived exosomes (SMSC-EXOs) and improves their miRNA profiles with an elevated proportion of beneficial miRNAs, while reducing harmful ones by pretreating SMSCs with Sr. Compared to untreated SMSC-EXOs, Sr-pretreated SMSC-derived exosomes (Sr-SMSC-EXOs) demonstrated superior therapeutic efficacy by mitigating chondrocyte ferroptosis and reducing osteoclast-mediated joint pain in TMJOA. Our results illustrate Alix's crucial role in Sr-triggered miRNA loading, identifying miR-143-3p as a key anti-TMJOA exosomal component. Interestingly, this system is specifically oriented towards synovium-derived stem cells. The insight into trace element-driven, site-specific miRNA selectively loading in SMSC-EXOs proposes a promising therapeutic enhancement strategy for TMJOA.
MicroRNAs/metabolism* ; Mesenchymal Stem Cells/drug effects* ; Osteoarthritis/drug therapy* ; Exosomes/drug effects* ; Strontium/pharmacology* ; Synovial Membrane/cytology* ; Humans ; Animals ; Temporomandibular Joint Disorders/therapy* ; Temporomandibular Joint

Objective:To analyze the expression of long-chain non-coding RNA U73166 in lung cancer tissues and its relationship with patients′ prognosis, and to explore the effects of silencing U73166 on proliferation and invasion of lung cancer H1299 cells and its regulatory mechanism. Methods:The expression level of U73166 in lung cancer tissues and normal tissues, as well as its correlation with lung cancer patients′ overall survival, were analyzed using the gene expression profiling interactive analysis (GEPIA) database. After culturing, H1299 cells were divided into a control group and a transfection group based on treatment conditions, and were transfected with 25 μmol/L of U73166 negative control and U73166 inhibitor, respectively. The effects of silencing U73166 on the relative expression of U73166 and microRNA-618 ( miR-618) genes in H1299 cells were assessed by real-time reverse transcription-PCR method. A cell counting kit-8 assay was used to evaluate the impact of silencing U73166 on the viability of H1299 cells. A transwell invasion assay was performed to detect the invasive ability of H1299 cells. The Linc2GO database and a dual-luciferase reporter assay were used to predict and verify the binding site between U73166 and miR-618. Western blotting was used to analyze the relative expression of phosphorylated Janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), and phosphorylated signal-transducing adaptor molecule 1 (p-STAM1) in the JAK2/STAT3 signaling pathway to evaluate the effects of silencing U73166 on this pathway in H1299 cells. Data were analyzed by an independent sample t test or one-way analysis of variance. Results:Analysis of the GEPIA database revealed that U73166 relative expression level in lung cancer tissues ( n=383) was significantly higher than that in normal tissues ( n=347) ( P<0.01). The overall survival of lung cancer patients with low U73166 expression [(245±2) months] was longer than that of patients with high U73166 expression [(167±2) months] ( P<0.05). The relative expression of U73166 were 7.81±0.99 in the control group and 1.01±0.26 in the transfection group, respectively, and the relative expression of miR-618 were 1.03±0.20 in the control group and 4.83±1.27 in the transfection group, respectively. Silencing U73166 significantly downregulated its expression ( t=6.66, P<0.01) and upregulated the relative expression of miR-618 ( t=2.96, P<0.01) in H1299 cells. After silencing U73166, the absorbance values of H1299 cells in the transfection group on days 2, 3, 4, and 5 (0.36±0.04, 0.74±0.05, 1.07±0.09, and 1.18±0.10) were significantly lower than those in the control group (0.55±0.03, 1.20±0.08, 1.63±0.07, and 1.90±0.07) ( P<0.05, 0.01). The number of invasive cells in the control and transfection groups were 52.03±6.08 and 19.92±3.78, respectively. There were significantly fewer invasive cells in the transfection group ( t=4.49, P<0.01). After transfection with wild-type U73166, the relative luciferase activity in the miR-618 group (0.32±0.05) was significantly lower than that in the miR-negtive control group (0.96±0.15) ( t=4.02, P<0.01). However, after transfection with mutant U73166, there was no statistically significant difference in relative luciferase activity between the miR-618 group (1.01±0.15) and the miR-negtive control group (1.03±0.11) ( t=0.09, P>0.05). The relative expression of p-JAK2, p-STAT3, and p-STAM1 proteins in the transfection group were 2.08±0.21, 1.36±0.20, and 0.55±0.12, respectively. These values were significantly lower than those in the control group (3.72?±?0.29, 5.56?±?0.19, and 4.38±0.17) (all P<0.01). Conclusions:U73166 is highly expressed in lung cancer tissues and lung cancer cells, and its expression is related to lung cancer patients′ overall survival. Silencing U73166 can target miR-618, which inhibits the proliferation and invasion of H1299 cells.

Objective Melatonin has been shown to have neuroprotective effects.This study is aimed at observing the effects of copper deposition on cognitive function in a toxic milk(TX)mouse model of Wilson disease(WD),and investigating the effects and mechanisms of action of Gandou Bushen Decoction(GDBSD)on melatonin synthesis and pineal function in the WD model mice.Methods A total of 30 homozygous TX mice were randomly assigned to 3 groups(n=10 in each group),including a WD group,a GDBSD group,and a dimercaptosuccinic acid(DMSA)group.A total of 10 DL mice were included in the normal control(NC)group.The structure and copper content of pineal gland tissues,oxidative stress and apoptosis-related markers,and serum melatonin levels were evaluated using hematoxylin-eosin(HE)staining,enzyme-linked immunosorbent assay(ELISA),flow cytometry,and Western blot.Results Compared with the NC group,the WD group exhibited decreased learning and cognitive abilities(P＜0.05),damaged pineal gland structure,increased copper content,reactive oxygen species(ROS)levels,and mitochondrial damage rate in the pineal gland(P＜0.01),altered levels of melatonin and oxidative stress-related markers(P＜0.05),upregulated expression levels of pro-apoptotic proteins Bax and Caspase-3,and decreased expression of the anti-apoptotic protein Bcl-2(P＜0.01).After treatment with GDBSD and DMSA,the SIRT3/FOXO3α signaling pathway was activated,the copper content in the pineal gland was reduced,and oxidative stress and apoptosis-related damages were improved,leading to an improvement in learning and memory abilities(P＜0.05).Conclusion GDBSD can alleviate cognitive impairments in WD mice caused by pineal gland copper deposition by inhibiting oxidative stress and apoptosis in the pineal gland.The underlying molecular mechanism is associated with the regulation of the SIRT3/FOXO3α signaling pathway.

Objective To investigate the status of occult hepatitis B virus infection(OBI)among blood donors in Quzhou,Zhejiang,and to analyze the mutation of S region in blood donors with anti-HBc+alone and non anti-HBc+alone.Methods The OBI samples were screened by ELISA and NAT;the HBV DNA was amplified and sequenced;20 anti-HBc+alone and 25 not anti-HBc+alone samples were obtained.Results The detection rate of OBI in Quzhou was 0.10%(155/161 045),and the positive rate of anti-HBc was 74.19%(115/155).The detection rate of OBI increased with the age of blood donors(P＜0.05),but was not related to gender.The positive rate of anti-HBc+alone was 22.58%(35/155),and that of not anti-HBc alone was 51.61%(80/155).Among the 45 OBI sequencing samples,the proportion of B and C geno-type was73.33%(33/45)and 20.00%(9/45),respectively.The mutation sites of blood donors in the anti-HBc+alone group were more than those in the not anti-HBc+alone group,and the mutation rates of S114T and V168A on MHR were significantly different(P＜0.05).Conclusion The genotype of OBI infection in Quzhou is mainly type B.The mutation sites of blood donors with anti-HBc+alone are higher than those with not anti-HBc+alone,which may be more suitable as one of the OBI screening indicators.

Objective: To retrospectively analyze the epidemiological characteristics of supernumerary teeth in patients aged 4-18 years old and the influencing factors on the selection of anesthesia methods, and to provide a reference for the selection of anesthesia plans for children and adolescents with supernumerary teeth. Methods: This study is a retrospective study approved by the Institutional Ethics Committee. Based on clinical electronic medical record system and cone beam CT (CBCT) data, a retrospective analysis was conducted on 2 210 patients 4-18 years of age who underwent supernumerary tooth extraction at the School of Stomatology, Fourth Military Medical University from August 2019 to July 2021. Inclusion criteria: ① Age 4-18 years old; and ② The American Society of Anesthesiologists (ASA) classifies anesthesia into grades I-II; and ③ Have complete oral and anesthesia case records and relevant imaging data. Exclusion criteria: ① Incomplete medical records or unclear imaging data; and ② Patients with ASA grade II or above. Patients’ gender and age, the number of supernumerary teeth, arch, location, orientation, eruption status, reason for appointment, anxiety level, degree of cooperation, anesthesia method, and other relevant information were collected and statistically analyzed. Results: A total of 1 865 eligible patients were included, with an average age of (8.9±3.2) years old. There were more male patients (71.37%, 1 331 cases) than female patients (28.63%, 534 cases) (P < 0.001), with a gender ratio of 2.49:1. The majority of supernumerary teeth were single (75.97%, 1 417/1 865), primarily located in the maxilla (97.2%, 1 812/1 865) and the anterior dental region (94.2%, 1 757/1 865), and in a centric position (77.3%, 1 442/1 865). The majority of patients with erupted supernumerary teeth were active in seeking treatment (97.67%, 335/343). Patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor, were mainly referred to the department for diagnosis, accounting for 94.38%(1 361/1 442) and 90.00% (72/80) (χ2=1 363.24, P < 0.001), respectively. Regarding anesthesia methods, the largest proportion of patients received nitrous oxide sedation-assisted local anesthesia or nerve block anesthesia, accounting for 38.07% (710/1 865), followed by local anesthesia, accounting for 35.23% (657/1 865). The proportion of patients receiving midazolam intravenous sedation with local anesthesia or nerve block anesthesia and general anesthesia was relatively small, accounting for 20.86% (389/1 865) and 5.84% (109/1 865). Patients 13-18 years of age with supernumerary teeth in the mandibular and posterior regions and without anxiety had the highest proportion of local anesthesia use (P < 0.001). In contrast, patients who had supernumerary teeth located at the base of the nose (50%), severe anxiety (94.12%), and poor cooperation (98.18%) had the highest proportion of general anesthesia use (P < 0.001). There was no significant difference (P = 0.35) in the incidence of intraoperative and postoperative complications after the extraction of supernumerary teeth. However, the proportion of anesthesia-related complications, such as dizziness, coughing, and respiratory depression, occurring in patients who received general anesthesia was higher than local anesthesia, accounting for 3.81% (P = 0.006). Conclusion There is a gender difference in the incidence of supernumerary teeth in patients 4-18 years of age, with a higher prevalence in males. The majority of supernumerary teeth are single and located in the maxillary anterior region, predominantly in a centric position. Patients whose teeth had erupted were more likely to seek medical treatment voluntarily, while patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor were more likely to be referred to the department. Patients with high levels of anxiety, poor cooperation, young age, multiple teeth, and high surgical difficulty were more inclined to choose general anesthesia.

Objective To investigate the clinical effects of sinus elevation surgery and implant restorationdue to in-sufficient bone massafter tooth extraction in patients with odontogenic maxillary sinusitis(OMS)and to provide a refer-ence for use in clinical practice.Methods This study was reviewed and approved by the Ethics Committee,and in-formed consent was obtained from the patients.Forty-five teeth were extracted from patients with OMS in the maxillary posterior area(the study group).Sinus elevation and implantation were performed due to insufficient bone height in the implant area 6-8 months after tooth extraction in the study group.Forty-eight teeth were extracted from patients without"OMS"in the maxillary posterior area(the control group),and sinus elevation and implantation were performed due to insufficient bone height in the implant area 6-8 months after tooth extraction inthe control group.In the study group,13 cases of discontinuous maxillary sinus floor bone and residual alveolar bone height of the maxillary sinus floor less than 4 mm were addressed with lateral wall sinus elevation,and the other 32 cases were addressed with crest-approach sinus elevation.In the control group,8 cases of residual alveolar bone height less than 4 mm in the maxillary sinus floor were addressed with lateral wall sinus,and the other 40 cases were addressed with crest approach sinus elevation.Restora-tions were placed 6 to 8 months after surgery.The patients were followed up 21 days,3 months,and 8 months after im-plantation and every 6 months after the placement of the restorations.The sinus bone gain(SBG),apical bone height(ABL)and marginal bone loss(MBL)were statistically analyzed 24 months after the restoration.Results The average preoperative mucosal thickness in the 45 patients in the study group was(1.556±0.693)mm,which was significantly larger than that in the control group(1.229±0.425)mm(P＜0.001).There were no perforations in either group.Twenty-four months after restoration,there was no significant difference in the SBG,ABH or MBL between the two groups(P＞0.05).Conclusion After the extraction of teeth from patients with OMS,the inflammation of the maxillary sinus de-creased,and the bone height and density in the edentulous area were restored to a certain degree.The effects of sinus floor lifting surgery and implant restoration do not differ between patients with and without OMS.