OBJECTIVE To investigate the mechanism by which esketamine improves postoperative cognitive impairment in rats with hip fracture based on the AMP-activated protein kinase （AMPK）/silencing information regulatory factor 1 （SIRT1）/ peroxisome proliferator activated-receptor-γ coactivator-1α （PGC-1α） signaling pathway. METHODS Rats with hip fracture surgery were assigned into model group， esketamine group （10 mg/kg）， inhibitor group （250 μg/mL AMPK inhibitor Compound C）， and esketamine+inhibitor group （10 mg/kg esketamine + 250 μg/mL Compound C）， and rats undergoing sham surgery were used as the control group， with 12 rats in each group. New object recognition and Barnes maze experiments were used to E-mail：448231@163.com evaluate cognitive function in rats. The levels of serum tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β），superoxide dismutase （SOD）， malondialdehyde （MDA）， gamma-aminobutyric acid （GABA）， dopamine （DA） and glutamate （Glu）， and the apoptosis of hippocampal neurons were detected. The pathological morphology of the hippocampal tissue and the ultrastructure of mitochondria were observed. The mRNA expression of B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X protein （Bax），the mRNA and protein expression of AMPK， SIRT1 and PGC-1α， as well as the expression of phosphorylated （p）-AMPK in hippocampal tissue， were detected. RESULTS Compared with the control group， the hippocampal neurons in the model group of rats were disordered， with more neurons necrotic and swollen mitochondria；the new object recognition index， the SOD， GABA， DA levels， Bcl-2， AMPK， SIRT1 and PGC-1α mRNA expression levels， and p-AMPK， SIRT1， PGC-1α protein expression levels were significantly reduced， while the latency and number of errors for locating unknown holes， the TNF-α， IL-1β， MDA and Glu levels， neuronal cell apoptosis rate， and Bax mRNA expression levels were significantly increased/prolonged （P＜0.05）. Compared with the model group， the esketamine group showed reduced pathological damage to the hippocampal tissue of rats， and the new object recognition index， the SOD， GABA and DA levels， the Bcl-2， AMPK， SIRT1 and PGC-1α mRNA expression levels， and p-AMPK， SIRT1， PGC-1α protein expression levels were significantly increased，while the latency and error frequency for locating unknown holes， TNF-α， IL-1β， MDA and Glu levels， neuronal cell apoptosis rate， and Bax mRNA expression levels were significantly decreased （P＜0.05）；the inhibitor group showed the opposite trend of changes in these indicators compared to the esketamine group （P＜0.05）.AMPK inhibitor could reverse the improvement effect of esketamine on the above indicators after hip fracture surgery in rats （P＜0.05）. CONCLUSIONS Esketamine may improve postoperative inflammatory response and oxidative stress levels in rats with hip fracture by activating the AMPK/SIRT1/PGC-1α signaling pathway， inhibiting neuronal cell apoptosis， improving mitochondrial structure， and promoting postoperative cognitive function recovery.

Objective:To construct a quality indicator system for pain nursing process in adult inpatients and to provide a scientific, objective and practical basis for the evaluation and management of pain nursing quality in clinical nursing.Methods:Based on the guidance framework of nursing procedures, a quality index system for pain nursing process in adult inpatients was preliminarily developed. Two rounds of expert questionnaires were conducted using the Delphi method, the analytic hierarchy process was used to analyze the weight of the indicators, and the semantic analysis of the indicator system was carried out with the nurses, and finally the quality indicator system for the pain nursing process in adult inpatients was constructed.Results:Both Delphi rounds attained 100% response rates, and the expert authority coefficients were 0.96 and 0.98 respectively, the familiarity coefficients were 0.95 and 0.98 respectively, both judgment basis coefficients were 0.98, and the Kendall′s coefficient of concordance were 0.187 and 0.451 respectively. The mean values of each indicator in semantic analysis ranged from 4.30 to 5.00, and the standard deviation was 0-1.06. The final constructed quality indicator system for the pain nursing process in adult inpatients included 5 primary-level indicators (pain assessment, pain nursing diagnosis, pain nursing plan, pain nursing measures, and pain nursing evaluation) and 28 secondary-level indicators.Conclusions:The quality indicator system for the pain nursing process is successfully constructed in adult inpatients, the method is scientific and reasonable, and the content is practical and reliable. It has guiding significance for evaluating the quality of pain nursing process for inpatients.

Objective:To investigate the effects of repetitive transcranial magnetic stimulation(rTMS) on the core symptoms, brain functional connectivity and activation in children with attention deficit hyperactivity disorder (ADHD) using functional near-infrared spectroscopy (fNIRS).Methods:From September 2022 to March 2024, a total of 35 children with ADHD were selected as research subjects and they were randomly divided into observation group ( n=17) and control group ( n=18). The control group received conventional rehabilitation therapy, while the observation group received rTMS therapy in addition to the conventional therapy. Both groups were treated every other day, with each course of treatment lasting four weeks, and a total of three courses of treatment were administered consecutively. The clinical symptoms of the children with ADHD were assessed using Swanson, Nolan, and Pelham-Ⅳ(SNAP-Ⅳ) before and after treatment. fNIRS was used to detect the relative concentration changes of oxyhemoglobin (HbO 2) and deoxyhemoglobin (HbR) in the prefrontal cortex under resting-state and Go/Nogo task conditions before and after treatment. Statistical analysis was performed using SPSS 26.0 software. Paired sample t-test were used for within-group comparisons, and independent sample t-test were used for between-group comparisons. Results:(1) After treatment, the scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the two groups were significantly lower than before treatment ( t=3.51-18.86, all P<0.05). The scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the observation group were significantly lower than those in the control group ( t=2.21, 2.03, 2.39, all P<0.05). (2) After treatment, the functional connectivity strength between all regions of interest in both groups was significantly higher than before treatment ( t=3.53-37.90, all P<0.05). The functional connectivity strength of the left dorsolateral prefrontal cortex (0.25±0.03, 0.21±0.03), right dorsolateral prefrontal cortex (0.12±0.02, 0.09±0.02), left medial prefrontal cortex (0.13±0.02, 0.10±0.01) and right medial prefrontal cortex (0.31±0.04, 0.24±0.06) in the observation group was significantly higher than those in the control group (all P<0.05). (3) In the Go/Nogo task, after treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex, right medial prefrontal cortex, left temporal lobe, and right temporal lobe in both groups were all higher than before treatment (all P<0.05). After treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex and right medial prefrontal cortex of the observation group were significantly higher than those of the control group (all P<0.05). Conclusion:rTMS therapy can improve the core symptoms of children with ADHD, which may be related to the strength of brain functional connectivity and activation of ADHD brain function by rTMS.

Objective Piperazine derivatives are a group of emerging psychoactive substances with excitatory and hallucinogenic effects on the central nervous system.This study established a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)screening method for the detection of 40 piperazine compounds in urine.Methods A 200 μL urine sample(spiked with an internal standard at 1 ng/mL)was subjected to liquid-liquid extraction with ethyl acetate.After nitrogen evaporation,the residue was redissolved in 200 μL methanol and injected for analysis.Separation was performed on a Waters Acquity UPLC? HSS T3 column(100 mm × 2.1 mm,1.8 μm).The mobile phase consisted of(A)20 mmol/L ammonium acetate buffer containing 0.1％formic acid and 5％acetonitrile,and(B)acetonitrile.Gradient elution was applied,and detection was carried out in multiple reaction monitoring(MRM)mode.Quantification was achieved using an internal standard calibration curve.Results The 40 piperazine substances demonstrated good linearity within the range of 1-50 ng/mL,with correlation coefficients of 0.995-0.998.The extraction recovery ranged from 51.51％to 104.1％.Intra-day precision was below 5％,while inter-day precision ranged from 1.61％to 10.17％.Accuracy was between-7.84％and 8.77％.The limits of detection were 0.2-1 ng/mL,and the limit of quantification was 1 ng/mL.Conclusion The proposed method requires only a small sample volume,exhibits high sensitivity,selectivity,and stability,and offers short run times.It is suitable for the qualitative and quantitative determination of piperazine derivatives in urine in forensic toxicology practice.

Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.

Objective:To investigate the effects of repetitive transcranial magnetic stimulation(rTMS) on the core symptoms, brain functional connectivity and activation in children with attention deficit hyperactivity disorder (ADHD) using functional near-infrared spectroscopy (fNIRS).Methods:From September 2022 to March 2024, a total of 35 children with ADHD were selected as research subjects and they were randomly divided into observation group ( n=17) and control group ( n=18). The control group received conventional rehabilitation therapy, while the observation group received rTMS therapy in addition to the conventional therapy. Both groups were treated every other day, with each course of treatment lasting four weeks, and a total of three courses of treatment were administered consecutively. The clinical symptoms of the children with ADHD were assessed using Swanson, Nolan, and Pelham-Ⅳ(SNAP-Ⅳ) before and after treatment. fNIRS was used to detect the relative concentration changes of oxyhemoglobin (HbO 2) and deoxyhemoglobin (HbR) in the prefrontal cortex under resting-state and Go/Nogo task conditions before and after treatment. Statistical analysis was performed using SPSS 26.0 software. Paired sample t-test were used for within-group comparisons, and independent sample t-test were used for between-group comparisons. Results:(1) After treatment, the scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the two groups were significantly lower than before treatment ( t=3.51-18.86, all P<0.05). The scores for inattention, hyperactivity/impulsivity and oppositional defiant behavior in the observation group were significantly lower than those in the control group ( t=2.21, 2.03, 2.39, all P<0.05). (2) After treatment, the functional connectivity strength between all regions of interest in both groups was significantly higher than before treatment ( t=3.53-37.90, all P<0.05). The functional connectivity strength of the left dorsolateral prefrontal cortex (0.25±0.03, 0.21±0.03), right dorsolateral prefrontal cortex (0.12±0.02, 0.09±0.02), left medial prefrontal cortex (0.13±0.02, 0.10±0.01) and right medial prefrontal cortex (0.31±0.04, 0.24±0.06) in the observation group was significantly higher than those in the control group (all P<0.05). (3) In the Go/Nogo task, after treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex, right medial prefrontal cortex, left temporal lobe, and right temporal lobe in both groups were all higher than before treatment (all P<0.05). After treatment, the average HbO 2 concentrations in the left dorsolateral prefrontal cortex, right dorsolateral prefrontal cortex, left medial prefrontal cortex and right medial prefrontal cortex of the observation group were significantly higher than those of the control group (all P<0.05). Conclusion:rTMS therapy can improve the core symptoms of children with ADHD, which may be related to the strength of brain functional connectivity and activation of ADHD brain function by rTMS.

Objective:To construct a quality indicator system for pain nursing process in adult inpatients and to provide a scientific, objective and practical basis for the evaluation and management of pain nursing quality in clinical nursing.Methods:Based on the guidance framework of nursing procedures, a quality index system for pain nursing process in adult inpatients was preliminarily developed. Two rounds of expert questionnaires were conducted using the Delphi method, the analytic hierarchy process was used to analyze the weight of the indicators, and the semantic analysis of the indicator system was carried out with the nurses, and finally the quality indicator system for the pain nursing process in adult inpatients was constructed.Results:Both Delphi rounds attained 100% response rates, and the expert authority coefficients were 0.96 and 0.98 respectively, the familiarity coefficients were 0.95 and 0.98 respectively, both judgment basis coefficients were 0.98, and the Kendall′s coefficient of concordance were 0.187 and 0.451 respectively. The mean values of each indicator in semantic analysis ranged from 4.30 to 5.00, and the standard deviation was 0-1.06. The final constructed quality indicator system for the pain nursing process in adult inpatients included 5 primary-level indicators (pain assessment, pain nursing diagnosis, pain nursing plan, pain nursing measures, and pain nursing evaluation) and 28 secondary-level indicators.Conclusions:The quality indicator system for the pain nursing process is successfully constructed in adult inpatients, the method is scientific and reasonable, and the content is practical and reliable. It has guiding significance for evaluating the quality of pain nursing process for inpatients.

Objective To analyze the symptomatic improvement effects of vitamin D in children with autism spectrum disorder(ASD)based on the microbiota-gut-brain axis.Methods Seventy-two children with ASD were randomly divided into an observation group and a control group,with 36 cases in each group.Three cases dropped out in the control group.The observation group received 1200 IU/day of vitamin D supplementation in addition to conventional rehabilitation training,while the control group received only conventional rehabilitation training.The intervention lasted for 12 weeks.Assessments were conducted before and after the intervention using the childhood autism rating scale(CARS),autism behavior checklist(ABC),and repetitive behavior scale-revised(RBS-R).Resting-state functional connectivity of the brain was measured using near-infrared functional imaging,and serum levels of 25(OH)D3,inflammatory cytokines,and gut microbiota were analyzed.The differences in these indicators before and after the intervention were compared between the two groups to evaluate clinical efficacy.Results The between-group differences in pre-and post-intervention changes showed that the observation group had significantly greater improvements than the control group in the following measures:CARS scores(-5.92±1.40 vs.-2.55±1.43),RBS-R scores(-5.99±1.01 vs.-3.10±1.47),resting-state brain functional connectivity(0.19±0.15 vs.0.10±0.18),serum 25(OH)D3 levels[(34.89±8.18)ng/mL vs.(0.68±6.73)ng/mL],serum interleukin-6(IL-6)levels[(-6.60±6.07)pg/mL vs.(-0.74±9.45)pg/mL],IL-1β levels[(-2.56±1.33)pg/mL vs.(-0.04±2.13)pg/mL],and tumor necrosis factor-α(TNF-α)levels[(-4.09±3.85)pg/mL vs.(0.21±4.05)pg/mL](P<0.05).Post-intervention,significant differences in gut microbial β-diversity were observed between the two groups(R2=0.030,P=0.040,Adonis).LEfSe analysis revealed that the observation group exhibited enrichment in Clostridia(LDA=4.747,P=0.003),Clostridiales(LDA=4.747,P=0.003),Clostridiaceae(LDA=3.476,P=0.001),Lachnospiraceae(LDA=4.709,P=0.004),Odoribacteraceae(LDA=3.458,P=0.027),Odoribacter(LDA=3.458,P=0.027),Burkholderiales(LDA=3.339,P=0.038),Firmicutes(LDA=4.764,P=0.003),and Betaproteobacteria(LDA=3.338,P=0.037).Conclusion Vitamin D supplementation can modulate gut microbial diversity in children with ASD,significantly influence the abundance of specific gut microbiota,reduce systemic inflammatory cytokines,enhance brain functional connectivity,and alleviate clinical symptoms of ASD.

Objective Piperazine derivatives are a group of emerging psychoactive substances with excitatory and hallucinogenic effects on the central nervous system.This study established a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)screening method for the detection of 40 piperazine compounds in urine.Methods A 200 μL urine sample(spiked with an internal standard at 1 ng/mL)was subjected to liquid-liquid extraction with ethyl acetate.After nitrogen evaporation,the residue was redissolved in 200 μL methanol and injected for analysis.Separation was performed on a Waters Acquity UPLC? HSS T3 column(100 mm × 2.1 mm,1.8 μm).The mobile phase consisted of(A)20 mmol/L ammonium acetate buffer containing 0.1％formic acid and 5％acetonitrile,and(B)acetonitrile.Gradient elution was applied,and detection was carried out in multiple reaction monitoring(MRM)mode.Quantification was achieved using an internal standard calibration curve.Results The 40 piperazine substances demonstrated good linearity within the range of 1-50 ng/mL,with correlation coefficients of 0.995-0.998.The extraction recovery ranged from 51.51％to 104.1％.Intra-day precision was below 5％,while inter-day precision ranged from 1.61％to 10.17％.Accuracy was between-7.84％and 8.77％.The limits of detection were 0.2-1 ng/mL,and the limit of quantification was 1 ng/mL.Conclusion The proposed method requires only a small sample volume,exhibits high sensitivity,selectivity,and stability,and offers short run times.It is suitable for the qualitative and quantitative determination of piperazine derivatives in urine in forensic toxicology practice.

Background and purpose:In 2016 the National Cancer Institute(NCI)decided stopping to use NCI-60 cell lines for drug screening,suggesting that tumor cell lines were losing their value as a tool for drug discovery and basic research.The reason for NCI-60 cells'retirement'was that the preclinical studies based on traditional cellular and animal models did not obtain the corresponding expected efficacy in clinical trials.Since the major cancer behaviors,such as proliferation and metastasis,are fundamentally altered with long-term culture,the tumor cell lines are not representative of the characteristics of cancer in patients.Currently,scientists hope to create a new cancer model that are derived from fresh patient samples and tagged with details about their clinical past.Our purpose was to create patient-derived breast cancer primary cell lines as new cancer model for drug screening and basic research.Methods:Breast cancer tissues were collected in the Department of Breast Surgery,Affiliated Hospital of Guizhou Medical University.The collection of tumor tissue samples was approved by the Ethics Committee of the Affiliated Hospital of Guizhou Medical University(approval number:2022 ethics No.313),and the collection and use of tumor tissues complied with the Declaration of Helsinki.The primary breast cancer cell lines were isolated from the patient's breast cancer tissues and cultured in BCMI medium.After the cells proliferated,the media were replaced with DEME medium.Cell line STR genotyping was done to determine cell-specific genetic markers and identification.Clone formation assay and transplantation assay were done to analyze the ability of breast cancer primary cell lines to form tumors.Results:We created 6 primary breast cancer cell lines.The 6 primary breast cancer cell lines from the patients were tagged with the definitively clinicopathological features,clinical diagnosis,therapeutic regimens,clinical effectiveness and prognostic outcomes.The STR genotyping assays identified the genetic markers and determined the identities of the 6 primary breast cancer cell lines.Clone formation assays and transplantation assay showed that the proliferative capacities of the patient-derived primary breast cancer cell lines were significantly greater compared with the conventional breast cancer cell lines.Conclusion:We created a panel of 6 patient-derived primary breast cancer cell lines as new cancer model for drug screening and basic research in breast cancer.