ObjectiveTo investigate the effect of interfering with long noncoding RNA （LncRNA） small nucleolar RNA host gene16 （SNHG16） on improving angiogenesis of ectopic endometrial stromal cells （EScs） in adenomyosis （AM） by targeting upregulation of miRNA （miR）-141-3p and inhibition of high mobility group box-1 protein （HMGB1）. MethodsThe expression levels of SNHG16， miR-141-3p and HMGB1 mRNA in endometrial tissues of 52 patients with adenomyosis （AM group） and 52 patients who needed hysterectomy due to cervical cancer or ovarian cancer （control group） were detected by quantitative reverse transcription polymerase chain reaction （qRT-PCR）. Y14 cells were divided into small hairpin RNA （shRNA） NC group， shRNA SNHG16 group， shRNA SNHG16+miR-141-3p inhibitor （inhibitor） group， shRNA SNHG16+inhibitor NC group and blank group. The targeting relationship between miR-141-3p and SNHG16 as well as HMGB1 was verified. The expression levels of SNHG16， miR-141-3p and HMGB1 mRNA in Y14 cells were detected by qRT-PCR. CCK-8 and Transwell assay were used to detect cell proliferation， invasion and migration. Microvascular density （MVD） was determined by immunofluorescence. The expressions of hypoxia-inducing factor α （HIF-1α）， cyclooxygenase-2 （Cox-2）， vascular endothelial growth factor （VEGF） and HMGB1 were detected by Western blot. ResultsThe expression of SNHG16 and HMGB1 mRNA in AM group was higher than that in control group， but the expression of miR-141-3p was lower than that in control group （P0.05）. The expression of SNHG16， HMGB1 mRNA， proliferation rate， migration， invasion number， MVD， expression of VEGF， HIF-1 α， Cox-2， and HMGB1 proteins in the shRNA SNHG16 group were lower than those in the blank group and shRNA NC group， while the expression of miR-141-3p was higher than that in the blank group and shRNA NC group （P0.05）. Inhibition of miR-141-3p reversed the improvement of EScs angiogenesis by interfering with SNHG16. ConclusionInterference with LncRNA SNHG16 improves EScs angiogenesis and inhibits proliferation， migration， and invasion of EScs by targeting upregulation of miR-141-3p and inhibition of HMGB1.

OBJECTIVE: To investigate effectiveness of simplified all-arthroscopic Broström technique in treatment of chronic lateral ankle instability in adolescents. METHODS: A clinical data of 21 adolescent patients with chronic lateral ankle instability, who met the selection criteria and were admitted between June 2023 and May 2024, was retrospectively analyzed. There were 18 males and 3 females with an average age of 16.0 years (range, 13-18 years). There were 9 cases of left ankle joint injury and 12 cases of right ankle joint injury. Anterior talofibular ligament (ATFL) injury was diagnosed by arthroscopy in all patients. There were 11 cases of cartilage injury, 5 cases of avulsion fractures, and 6 cases of ankle impingement syndrome. The time from first sprain to operation ranged from 3-60 months (mean, 12.0 months). The ATFL was repaired and the ankle joint stability was restored by simplified all-arthroscopic Broström technique. Visual analogue scale (VAS) score, Tegner score, American Orthopaedic Foot and Ankle Society (AOFAS) score, Karlsson ankle function scale (KAFS) score, Foot and Ankle Outcome Score (FAOS) were used to evaluate ankle pain and function. MRI was used to evaluate the ligament healing. RESULTS: All patients were followed up 8-15 months (mean, 12.6 months). After operation, 1 patient suffered from superficial peroneal nerve injury, 1 patient developed anterior scar impingement on the ankle, 2 patients had superficial wound infection, and 1 patient suffered from sprain again. The VAS score, Tenger score, AOFAS score, KAFS score, and FAOS score significantly improved when compared with the preoperative scores ( P<0.05). MRI examination showed the ligament healing and good tension. CONCLUSION For adolescent patients with chronic lateral ankle instability, using simplified all-arthroscopic Broström technique to repair ATFL can effectively alleviate ankle pain, improve stability, and achieve good effectiveness.
Humans ; Joint Instability/surgery* ; Male ; Adolescent ; Female ; Arthroscopy/methods* ; Ankle Joint/physiopathology* ; Retrospective Studies ; Ankle Injuries/surgery* ; Lateral Ligament, Ankle/injuries* ; Treatment Outcome ; Chronic Disease ; Range of Motion, Articular

Objective To develop a rapid and accurate ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method for the detection of etomidate,metomidate,propoxate,and isopropoxate in human hair.Methods Hair samples containing etomidate,metomidate,propoxate,and isopropoxate were extracted with methanol containing the internal standard orthoxine,filtered with a 0.22 μm organic filter membrane and detected vio UPLC-MS/MS.All components were separated by using a gradient elution procedure consisting of 0.01％formic acid(1 mmol/L ammonium acetate)and acetonitrile.Positive electrospray ionization was performed using multiple reaction monitoring(MRM)mode.Results The linear relationships of etomidate,metomidate,propoxate,and isopropoxate were good in the range of 0.01～1 ng/mg(r ≥ 0.997 9),with recovery rates ranging from 87.9％to 101.5％.The accuracy was between 80.0％and 110.0％.The intra-day and inter-day relative standard deviations(RSD)were 2.9％～9.6％and 3.6％～19.9％.Conclusion This method is easy to operate and has high recovery efficiencies.It is sufficiently simple and sensitive to be applied to detect etomidate,metomidate,propoxate,and isopropoxate in hair.

Objective To investigate the regulation of synaptic plasticity by cannabinoid receptor 1(CB1R)and its effects on autism spectrum disorder(ASD)-like behavior.Methods CB1R-knockout(KO)mice and valproic acid(VPA)-induced ASD model mice(VPA mice)were used as study subjects.Behavioral experiments were used to assess the effects of CB1R on ASD-like behavior in mice,neuronal structural integrity and dendritic density were detected by microtubule-associated protein 2(MAP2)staining experiments,and the expression of synapse-associated proteins was detected by Western blot,to assess the effects of CB1R on synaptic plasticity.Results Behavioral result showed that VPA mice demonstrated significant ASD-like behavior,while CB1R-/-mice spent a significantly smaller proportion of residence time in the central region of the open field(P＜0.0001),showed significant increases in the number of marbles buried and self-grooming time(P＜0.01),significantly less time spent socializing with unfamiliar mice 2 and exploring unfamiliar objects(P＜0.001),and significantly more time exploring old objects(P＜0.05).The relative dwelling time was significantly reduced in CB1R+/-mice(P＜0.001),and the number of marbles buried and self-grooming time were significantly increased(P＜0.05).Synaptic plasticity assays revealed significant synaptic plasticity impairment in VPA mice.Hippocampal MAP2-positive neuron densities were significantly reduced in CB1R-/-and CB1R+/-mice,and expression levels of synapsin-1 were significantly increased(P＜0.05).Conclusions CB1R KO leads to ASD-like behavior such as anxiety and repetitive stereotyped behavior,social and cognitive impairments,as well as neuronal damage,dendritic dysplasia and disrupted synaptic protein expression in mice,suggesting that CB1R is involved in regulating synaptic plasticity as a pathological mechanism for the development of ASD-like behavior.

Objective To develop a rapid and accurate ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method for the detection of etomidate,metomidate,propoxate,and isopropoxate in human hair.Methods Hair samples containing etomidate,metomidate,propoxate,and isopropoxate were extracted with methanol containing the internal standard orthoxine,filtered with a 0.22 μm organic filter membrane and detected vio UPLC-MS/MS.All components were separated by using a gradient elution procedure consisting of 0.01％formic acid(1 mmol/L ammonium acetate)and acetonitrile.Positive electrospray ionization was performed using multiple reaction monitoring(MRM)mode.Results The linear relationships of etomidate,metomidate,propoxate,and isopropoxate were good in the range of 0.01～1 ng/mg(r ≥ 0.997 9),with recovery rates ranging from 87.9％to 101.5％.The accuracy was between 80.0％and 110.0％.The intra-day and inter-day relative standard deviations(RSD)were 2.9％～9.6％and 3.6％～19.9％.Conclusion This method is easy to operate and has high recovery efficiencies.It is sufficiently simple and sensitive to be applied to detect etomidate,metomidate,propoxate,and isopropoxate in hair.

Objective To investigate the regulation of synaptic plasticity by cannabinoid receptor 1(CB1R)and its effects on autism spectrum disorder(ASD)-like behavior.Methods CB1R-knockout(KO)mice and valproic acid(VPA)-induced ASD model mice(VPA mice)were used as study subjects.Behavioral experiments were used to assess the effects of CB1R on ASD-like behavior in mice,neuronal structural integrity and dendritic density were detected by microtubule-associated protein 2(MAP2)staining experiments,and the expression of synapse-associated proteins was detected by Western blot,to assess the effects of CB1R on synaptic plasticity.Results Behavioral result showed that VPA mice demonstrated significant ASD-like behavior,while CB1R-/-mice spent a significantly smaller proportion of residence time in the central region of the open field(P＜0.0001),showed significant increases in the number of marbles buried and self-grooming time(P＜0.01),significantly less time spent socializing with unfamiliar mice 2 and exploring unfamiliar objects(P＜0.001),and significantly more time exploring old objects(P＜0.05).The relative dwelling time was significantly reduced in CB1R+/-mice(P＜0.001),and the number of marbles buried and self-grooming time were significantly increased(P＜0.05).Synaptic plasticity assays revealed significant synaptic plasticity impairment in VPA mice.Hippocampal MAP2-positive neuron densities were significantly reduced in CB1R-/-and CB1R+/-mice,and expression levels of synapsin-1 were significantly increased(P＜0.05).Conclusions CB1R KO leads to ASD-like behavior such as anxiety and repetitive stereotyped behavior,social and cognitive impairments,as well as neuronal damage,dendritic dysplasia and disrupted synaptic protein expression in mice,suggesting that CB1R is involved in regulating synaptic plasticity as a pathological mechanism for the development of ASD-like behavior.

【Objective】 To explore the relationship between the methylation level of CNR1 and autism spectrum disorder (ASD), in order to provide a theoretical basis for the etiology of ASD. 【Methods】 A case-control study was conducted, recruiting 30 children with ASD from the Child Development and Behavior Research Center of Harbin Medical University and a rehabilitation facility, and 30 matched typically developed children from June 2017 to December 2018. The methylation levels of CNR1 in peripheral blood were measured by the Agena MassArray^® Mass Spectrometry System. A univariate conditional Logistic regression model was used to analyze the potential association between the methylation level of CNR1 and the risk of ASD with adjustment for age, BMI, body fat percentage and body fat. The correlations between the methylation level of CNR1 and the score of Social Responsiveness Scale (SRS) were evaluated by Pearson/Spearman correlation analysis. 【Results】 The methylation levels of the average methylation (t=2.224), CpG_3.4 (Z=2.187), CpG_9.10.11 (t=2.308), and CpG_28.29 (t=2.943) of the CNR1 promoter region in ASD children were significantly higher than controls (P<0.05). The methylation levels of the average methylation (OR=1.117, 95%CI: 1.003 - 1.245), CpG_9.10.11 (OR= 1.072, 95%CI:1.006 - 1.142), and CpG_28.29 (OR=1.078, 95%CI: 1.018 - 1.141) of the CNR1 promoter region were positively correlated with the risk of ASD (P<0.05). The methylation level of CpG_28.29 in ASD children was positively correlated with the scores of social motivation in SRS (r=0.421, P<0.05). 【Conclusions】 The methylation levels of CNR1 in peripheral blood are abnormal in ASD children and might be correlated with the risk of ASD and social function. The underlying mechanism needs to be further explored.

Objective: Autism spectrum disorder (ASD) is a neurodevelopmental disorder with onset in infancy. Early intervention is critical to improve the prognosis for these children. E-health interventions have tremendous potential. This review aimed to determine the status and effectiveness of family interventions for parents of children aged 0–6 years with ASD in the context of e-health. Methods: The review methodology was guided by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews. PubMed, Web of Science, and China National Knowledge Infrastructure were searched from inception to June 2022. The searches were limited to children with ASD of the age range between 0 and 6 years. We collated the available information and used descriptive statistics to analyze the synthesized data. Results: Our initial search identified 3,672 articles, of which 30 studies met the inclusion criteria. The 30 articles selected were released between 2012 and 2022. All articles are in English. Most articles reviewed were from high-income countries (27/30, 90.0%), especially from the United States (16/30, 53.3%). Four major themes emerged from the 30 studies that matched the inclusion criteria, as follows: 1) type of e-health interventions, 2) duration of interventions, 3) clinical aspects of e-health interventions, and 4) evidence for intervention effectiveness, looking into the positive, negative, and mixed findings of previous studies. Conclusion These findings suggest that a wide variety of e-health interventions may actually help support both children with ASD aged 0–6 years and their parents.

Objective:To explore the relationship between endocannabinoid (eCB) and its metabolic enzymes and severity of autism spectrum disorder (ASD), and to provide a theoretical basis for the study of the etiology and pathogenesis of ASD.Methods:A case-control study was conducted to collect 58 ASD children who underwent rehabilitation training at the Children's Developmental Behavior Research Center of Harbin Medical University and the provincial autism rehabilitation facility from December 2017 to December 2018 as the ASD group.According to the principle of gender and age 1∶1 matching, 58 normal children were selected as control group in Heilongjiang Province.Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and real-time quantitative PCR (RT-qPCR) were used to detect eCB of ASD group and control group, including anandamide (AEA), 2-arachidonoylglycerol (2-AG), palmitoylethanolamide (PEA), oleoylethanolamide (OEA) and its metabolic enzymes: n-acylphosphatidylethanolamine-specific phospholipase D (NAPE-PLD), fatty acid amide hydrolase (FAAH), monoacylglycerol lipase (MAGL) and diacylglycerol lipase (DAGL) mRNA expression levels.Pearson correlation was used to analyze the level of eCB and ASD children's severity.Results:The levels of AEA, OEA and PEA in ASD children ((10.10±2.6)nmol/L, (24.30±5.60)nmol/L, (15.92±2.28)nmol/L) were lower than those in the control group ((13.46±3.04)nmol/L, (27.85± 6.89)nmol/L, (17.87±2.67)nmol/L, t=-6.612, -3.99, -4.779, P<0.01). The expression levels of FAAH and DAGL mRNA in ASD children were significantly higher than those in the control group, and the difference was statistically significant( t=2.423, 3.840, P<0.05), while NAPE-PLD and MAGL mRNA levels were not significantly different between the two groups ( t=0.024, 0.885, P>0.05). The level of PEA in the ASD group was negatively correlated with the total score of the autism behavior checklist (ABC) ( r=-0.288, P<0.05). Conclusion:There may be metabolic abnormalities in eCB and its metabolic enzymes in ASD children, and the level of eCB is related with the severity of ASD.

Objective: To investigate the level of vitamin B12 in children with autism spectrum disorder(ASD), and provide a theoretical basis for early detection and drug treatment of ASD. Methods: A total of 89 ASD cases and 89 matched controls were collected. The levels of urinary methylmalonic acid (MMA) and serum vitamin B12, Transcobalamin Ⅱ (TCN2) were determined by enzyme-linked immunosorbent assay (ELISA). TCN2 gene rs1801198 was genotyped by SNaPshot. Results: The serum levels of vitamin B12 and TCN2 in children with ASD [(369.08±131.88)pmol/L, (1.56±0.16)ng/mL] were significantly lower than those in the control group[(485.16±200.33)pmol/L, (1.71±0.17)ng/mL](t=-5.47, -5.92, P<0.05). The level of MMA in urine of ASD children [(758.97±106.96) ng/mL] was significantly higher than that in the control group[(693.66±121.72)ng/mL](t=3.94, P<0.05); The genetic polymorphism of rs1801198 locus was not associated with the risk of ASD(P>0.05), and there was no significant correlation with serum TCN2 level(F=1.16, P>0.05). Conclusion ASD children are at a potential deficiency of vitamin B12 and should strengthen their nutritional interventions while conducting ASD interventions.