BACKGROUND:Previous studies have found that neuronal damage caused by continuous excessive fluoride exposure is related to Ca2+overload,but the mechanism of Ca2+flow conversion between intracellular calcium stores and cell apoptosis damage is still unclear.OBJECTIVE:To investigate the effect of fluoride exposure on Ca2+transport channel proteins and apoptosis levels in the mitochondria-associated endoplasmic reticulum membrane of primary cultured neural cells.METHODS:Primary nerve cells of neonatal SD rats were cultured in vitro and identified by immunofluorescence staining with neuronal nucleus-specific antibody up to day 7.The nerve cells were divided into control group(containing 0 mmol/L sodium fluoride),low fluoride group(containing 0.5 mmol/L sodium fluoride),and high fluoride group(containing 1 mmol/L sodium fluoride).The cell morphological changes were observed by light microscope 24 hours after fluorine exposure.The expression levels of apoptosis-related protein BAX/BCL-2 and calcium transfer-related pathways VDAC1,GRP 75,and IP3R were detected using western blot assay.The expression levels of VDAC1,GRP 75,and IP3R mRNA were detected by RT-PCR.Ca2+levels were detected by Rhood-2AM Ca2+probe.Mitochondrial membrane potential detection kit was used to detect the change in mitochondrial membrane potential.The level of apoptosis was determined by flow cytometry and TUNEL staining.RESULTS AND CONCLUSION:(1)The purity of neurons cultured on day 7 had been determined to be over 90％,with few impurities,good growth status,and tight cell network connections,meeting the requirements of subsequent experiments.(2)Compared with the control group,growth of neural cell clusters in the low-fluoride group and the high-fluoride group increased;the processes were broken;the cell body was rounded,and the connection network between cells was destroyed.Compared with the low-fluoride group,the cell damage changes in the high-fluoride group were more obvious.(3)Compared with the control group,the protein expressions of VDAC1,GRP75,and IP3R were increased in the low-fluoride group and the high-fluoride group(P＜0.05),and the ratio of apoptosis-related protein BAX/BCL-2 was increased(P＜0.05).Compared with the control group,the expression of VDAC1 and GRP75 mRNA in the low-fluoride group was significantly increased(P＜0.05);the expression levels of VDAC1,GRP75,and IP3R mRNA in the high-fluoride group were significantly increased(P＜0.01).(4)The level of cell apoptosis increased significantly after fluoride exposure,and the high-fluoride group was significantly higher than the control and low-fluoride groups(P＜0.01).(5)After fluoride exposure,the concentration of mitochondrial Ca2+in nerve cells increased significantly(P＜0.05),the mitochondrial membrane potential decreased(P＜0.01),and the degree of damage in the high-fluoride group was more obvious(P＜0.05).The results show that fluoride exposure impairs the morphological structure of primary neural cells,resulting in upregulation of Ca2+transfer pathway protein expression between the endoplasmic reticulum and mitochondria,mitochondrial Ca2+overload,mitochondrial damage,and increased levels of apoptosis.

BACKGROUND:Previous studies have found that neuronal damage caused by continuous excessive fluoride exposure is related to Ca2+overload,but the mechanism of Ca2+flow conversion between intracellular calcium stores and cell apoptosis damage is still unclear.OBJECTIVE:To investigate the effect of fluoride exposure on Ca2+transport channel proteins and apoptosis levels in the mitochondria-associated endoplasmic reticulum membrane of primary cultured neural cells.METHODS:Primary nerve cells of neonatal SD rats were cultured in vitro and identified by immunofluorescence staining with neuronal nucleus-specific antibody up to day 7.The nerve cells were divided into control group(containing 0 mmol/L sodium fluoride),low fluoride group(containing 0.5 mmol/L sodium fluoride),and high fluoride group(containing 1 mmol/L sodium fluoride).The cell morphological changes were observed by light microscope 24 hours after fluorine exposure.The expression levels of apoptosis-related protein BAX/BCL-2 and calcium transfer-related pathways VDAC1,GRP 75,and IP3R were detected using western blot assay.The expression levels of VDAC1,GRP 75,and IP3R mRNA were detected by RT-PCR.Ca2+levels were detected by Rhood-2AM Ca2+probe.Mitochondrial membrane potential detection kit was used to detect the change in mitochondrial membrane potential.The level of apoptosis was determined by flow cytometry and TUNEL staining.RESULTS AND CONCLUSION:(1)The purity of neurons cultured on day 7 had been determined to be over 90％,with few impurities,good growth status,and tight cell network connections,meeting the requirements of subsequent experiments.(2)Compared with the control group,growth of neural cell clusters in the low-fluoride group and the high-fluoride group increased;the processes were broken;the cell body was rounded,and the connection network between cells was destroyed.Compared with the low-fluoride group,the cell damage changes in the high-fluoride group were more obvious.(3)Compared with the control group,the protein expressions of VDAC1,GRP75,and IP3R were increased in the low-fluoride group and the high-fluoride group(P＜0.05),and the ratio of apoptosis-related protein BAX/BCL-2 was increased(P＜0.05).Compared with the control group,the expression of VDAC1 and GRP75 mRNA in the low-fluoride group was significantly increased(P＜0.05);the expression levels of VDAC1,GRP75,and IP3R mRNA in the high-fluoride group were significantly increased(P＜0.01).(4)The level of cell apoptosis increased significantly after fluoride exposure,and the high-fluoride group was significantly higher than the control and low-fluoride groups(P＜0.01).(5)After fluoride exposure,the concentration of mitochondrial Ca2+in nerve cells increased significantly(P＜0.05),the mitochondrial membrane potential decreased(P＜0.01),and the degree of damage in the high-fluoride group was more obvious(P＜0.05).The results show that fluoride exposure impairs the morphological structure of primary neural cells,resulting in upregulation of Ca2+transfer pathway protein expression between the endoplasmic reticulum and mitochondria,mitochondrial Ca2+overload,mitochondrial damage,and increased levels of apoptosis.

ObjectiveTo investigate the liver histopathological features of HBeAg-negative patients in the indeterminate phase of low-viral-load chronic hepatitis B virus （HBV） infection. MethodsA total of 271 patients with low-viral-load HBeAg-negative chronic HBV infection who underwent liver biopsy in Department of Infectious Diseases， Affiliated Hospital of Yan’an University， from September 2013 to June 2021 were enrolled as subjects， and the degree of liver injury was compared between patients based on age， sex， presence or absence of the family history of hepatitis B， HBsAg， and alanine aminotransferase （ALT） level. The chi-square test was used for comparison of categorical data between two groups. ResultsAmong the 271 patients with HBeAg-negative chronic HBV infection， 86 patients （31.73%） grade≥A2 liver inflammatory activity， 72 （26.57%） had a liver fibrosis stage of ive， and 112 （41.33%） had moderate or severe liver histological injury. The proportion of patients with grade≥A2 liver inflammatory activity in the patients with ALT>20 U/L was significantly higher than that in the patients with ALT≤20 U/L （χ²=3.938， P=0.047）. There were no significant differences in the proportion of patients with grade≥A2 liver inflammatory activity between the patients with different ages， sexes， family history of hepatitis B， HBsAg levels （all P>0.05），there were no significant differences in the proportion of patients with a liver fibrosis stage of ≥F2 between the patients with different ages， sexes， family history of hepatitis B， HBsAg， and ALT levels （all P>0.05）， and the stratified analysis of patients aged≤30 years and patients without the family history of hepatitis B showed no statistical significance between groups （all P>0.05）. There was no significant difference in the degree of liver histological injury between the patients with different ages， sexes， family history of hepatitis B， HBsAg， and ALT levels （all P>0.05）. ConclusionSignificant liver injury is observed in more than 40% of the patients with low-viral-load HBeAg-negative chronic HBV infection， and there is no significant difference in the degree of liver histological injury between the patients with different ages， sexes， family history of hepatitis B， HBsAg， and ALT levels. Even for the patients aged≤30 years who deny the family history of hepatitis B， there is still a considerable proportion of patients with liver injury， which should be taken seriously by clinicians.

ObjectiveTo elucidate the critical role of rehabilitation medical records (including electronic records) in rehabilitation medicine's clinical practice and management, comprehensively analyzed the structure, core content and data standards of rehabilitation medical records, to develop a standardized medical record data architecture and core dataset suitable for rehabilitation medicine and to explore the application of rehabilitation data in performance evaluation and payment. MethodsBased on the regulatory documents Basic Specifications for Medical Record Writing and Basic Specifications for Electronic Medical Records (Trial) issued by National Health Commission of China, and referencing the World Health Organization (WHO) Family of International Classifications (WHO-FICs) classifications, International Classification of Diseases (ICD-10/ICD-11), International Classification of Functioning, Disability and Health (ICF), and International Classification of Health Interventions (ICHI Beta-3), this study constructed the data architecture, core content and data standards for rehabilitation medical records. Furthermore, it explored the application of rehabilitation record summary sheets (home page) data in rehabilitation medical statistics and payment methods, including Diagnosis-related Groups (DRG), Diagnosis-Intervention Packet (DIP) and Case Mix Index. ResultsThis study proposed a systematic standard framework for rehabilitation medical records, covering key components such as patient demographics, rehabilitation diagnosis, functional assessment, rehabilitation treatment prescriptions, progress evaluations and discharge summaries. The research analyzed the systematic application methods and data standards of ICD-10/ICD-11, ICF and ICHI Beta-3 in the fields of medical record terminology, coding and assessment. Constructing a standardized data structure and data standards for rehabilitation medical records can significantly improve the quality of data reporting based on the medical record summary sheet, thereby enhancing the quality control of rehabilitation services, effectively supporting the optimization of rehabilitation medical insurance payment mechanisms, and contributing to the establishment of rehabilitation medical performance evaluation and payment based on DRG and DIP. ConclusionStructured rehabilitation records and data standardization are crucial tools for quality control in rehabilitation. Systematically applying the three reference classifications of the WHO-FICs, and aligning with national medical record and electronic health record specifications, facilitate the development of a standardized rehabilitation record architecture and core dataset. Standardizing rehabilitation care pathways based on the ICF methodology, and developing ICF- and ICD-11-based rehabilitation assessment tools, auxiliary diagnostic and therapeutic systems, and supporting terminology and coding systems, can effectively enhance the quality of rehabilitation records and enable interoperability and sharing of rehabilitation data with other medical data, ultimately improving the quality and safety of rehabilitation services.

AIM To explore the clinical effects of Tongfu Jiangzhuo Formula on ICU delirium patients with Internal Accumulation of Turbid Toxins.METHODS Based on Turbid Toxins theory,two hundred and ten patients were randomly assigned into control group(105 cases)for intervention of conventional treatment,and observation group(105 cases)for intervention of both Tongfu Jiangzhuo Formula and conventional treatment.The changes in clinical effects,delirium scores(CAM-ICU,ICDSC),TCM syndrome score,inflammatory factors(IL-1β,IL-6,IL-8),Raf-1,MEK-2,ERK-1 mRNA and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed decreased delirium scores,TCM syndrome score,inflammatory factors and Raf-1,MEK-2,ERK-1 mRNA(P＜0.05),especially for the observation group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with ICU delirium due to Internal Accumulation of Turbid Toxins,Tongfu Jiangzhuo Formula can safely and effectively improve symptoms,and reduce inflammation levels and Raf-1,MEK-2,ERK-1 mRNA expressions.

Objective:To retrospectively analyse the clinical data of patients after NUSS procedure for pectus excavatum, and summary of surgical techniques for NUSS bar removal.Methods:Retrospectively collected the clinical data of 276 patients undergoing NUSS bar removal from January 2024 to September 2024 in Beijing Children's Hospital affiliated to Capital Medical University. The age of the patients ranged from 6 to 20 years old, 211 males and 65 females. The average time the bar was in place in the body was 36 months.Results:All 276 patients successfully completed the NUSS bar removal. The average operative time was 22.6 min, with an average blood loss of 3.3 ml. 90 patients with bone scabs, 104 patients with wire breakage were successfully removed. 2 cases of postoperative wound infection, no other intraoperative and postoperative complications. The average hospitalization time after surgery was 1.2 days. Follow up for 3 months after surgery, and no abnormalities were found on the chest X-ray.Conclusion:Mastering the surgical techniques for pectus excavatum bar removal enhances the safety and efficiency of the procedure. It effectively reduces the incidence of intraoperative and postoperative complications, shortens operative time, and alleviates postoperative pain in patients.

Objective:To analyze and identify the expression profiles of oxidative stress-related genes and circular RNAs(circRNAs)in ricin toxin(RT)-induced pyroptosis of mouse mononuclear macrophages(RAW264.7)using transcriptome sequencing and bioinformatics technology,and to preliminarily analyze their potential functions.Methods:The macrophages(RAW264.7 cells)were treated with RT to establish a cell pyroptosis model and divided into control group,40 μg·L-1 RT group,and 80 ng/mL RT group.Transmission electron microscope(TEM)was used to observe the morphology of the RAW264.7 cells in various groups;Western blotting method was used to detect the expression levels of the pyroptosis pathway-related proteins in the cells in various groups;80 μg·L-1 RT was selected for subsequent experiments.Transcriptome sequencing(RNA-Seq)was performed to obtain the circRNA and mRNA expression profiles in the RAW264.7 cells in control group and RT-treated group,followed by bioinformatics analysis.Results:Compared with control group,the cells in 40 and 80 μg·L-1 RT groups underwent morphological changes;the cells in RT groups showed obvious pyroptosis-like morphological changes,characterized by significant swelling of dying cells and the appearance of characteristic large bubbles on the plasma membrane.Compared with control group,the expression level of gasdermin DN-terminal fragment(GSDMD-N)protein in 40 and 80 μg·L-1 RT groups was increased(P<0.05);compared with 40 μg·L-1 RT group,the expression level of GSDMD-N protein in the cells in 80 μg·L-1 RT group was increased(P<0.05);therefore,the subsequent experiments used the RT concentration of 80 μg·L-1.A total of 2930 differentially expressed messenger RNAs(mRNAs)and 24 differentially expressed circRNAs were identified.The constructed circRNA-microRNA(miRNA)-mRNA competing endogenous RNA(ceRNA)regulatory network consisted of 7 circRNAs,12 miRNAs and 13 mRNAs.Gene Ontology(GO)functional enrichment analysis showed that in biological process(BP),the functions regulated by differentially expressed genes mainly included immune response and oxidative stress response;in cellular component(CC),differentially expressed genes were mainly localized to the external side of plasma membrane and cell leading edge;in molecular function(MF),they were mainly involved in transporter transmembrane activity and hormone receptor binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that differentially expressed genes were mainly enriched in Toll-like receptor signaling pathway,Forkhead box O(FoxO)signaling pathway and nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB)signaling pathway.In the protein-protein interaction(PPI)network,the top 10 hub genes with the highest connectivity were screened by CytoHubba,including matrix metalloproteinase 9(MMP9),superoxide dismutase 2(SOD2),and v-src sarcoma viral oncogene homolog(Src).Conclusion:The expression profiles of oxidative stress-related genes and circRNAs in RAW264.7 cells are altered after RT treatment.The screened differentially expressed circRNAs and mRNAs may serve as potential targets to regulate RT-induced pyroptosis in RAW264.7 cells through oxidative stress pathways.

Objective:To investigate the effects of brief mindfulness-based stress reduction(MBSR)on preoperative anxiety in pa-tients undergoing painless gastrointestinal endoscopy.Methods:We enrolled 100 patients scheduled to undergo elective painless gas-trointestinal endoscopy at The First Affiliated Hospital of Zhengzhou University from September 2024 to April 2025.The inclusion cri-teria were:age,18-60 years;body mass index,18.0-28.0 kg/m2;American Society of Anesthesiologists physical status,class Ⅰ orⅡ;and no gender restriction.The patients were assigned to experimental group(n=50)or control group(n=50)using a random num-ber table.A dedicated nursing team implemented the brief MBSR protocol.At 30 minutes before endoscopy,both groups underwent anxiety assessment using the Amsterdam Preoperative Anxiety and Information Scale(APAIS).All the patients received routine preop-erative education.Guided by the nurses,the experimental group received the brief MBSR intervention consisting of mindful body scan-ning,mindful breathing,and mindful music listening,for 12 minutes each at 30 and 15 minutes before the procedure.We recorded the APAIS score,bispectral index(BIS),heart rate(HR),systolic blood pressure(SBP),diastolic blood pressure(DBP),and mean arterial pressure(MAP)at 30 minutes before the procedure(T0),after brief MBSR(T1),and immediately before anesthesia induction(T2);the length of stay in the post-anesthesia care unit(PACU)and postoperative adverse reactions;and the APAIS score and degree of sat-isfaction of patients at discharge from the PACU(T3).Results:Com-(all P<0.05).However,no significant differences were observed be-pared with the control group,the experimental group exhibited sig-nificantly lower APAIS scores,significantly reduced BIS values,and significantly lower HR values at T1 and T2 and a significantly lower APAIS score and a significantly higher degree of satisfaction at T3 tween the groups in SBP,DBP,MAP,postoperative adverse events,or PACU length of stay at any time point(all P>0.05).Conclusion:Brief MBSR is an effective non-pharmacological intervention to cope with perioperative negative emotions in patients undergoing pain-less gastrointestinal endoscopy,which can alleviate preoperative anxiety,reduce electroencephalographic arousal,and improve patient satisfaction.

OBJECTIVE To investigate the roles and mechanisms of γ-bungarotoxin(γ-BGT)in inducing respiratory distress in mice.METHODS Six male Kunming mice were selected and anesthe-tized before tracheal intubation and respiratory recording.After stabilizing respiration,the mice were intraperitoneally injected with γ-BGT at a dose of 6 mg·kg-1.Once a decrease in respiratory frequency was observed,the mice were intravenously injected with nikethamide at a dose of 12.5 mg·kg-1.Respi-ratory frequency was monitored using the BL420 signal acquisition and processing system.Male Kunming mice were randomly divided into the normal control group(saline,ip),γ-BGT group(6 mg·kg-1,ip),and γ-BGT+nikethamide group(γ-BGT 6 mg·kg-1,ip,followed by nikethamide 12.5 mg·kg-1,ip,when shal-low breathing and enhanced abdominal respiration were observed).The levels of Glu and GABA in the medulla oblongata were measured using ELISA.The protein expression levels of GAD65 and GAD67 in the medulla oblongata were determined by Western blotting.Primary mouse medullary neurons were cultured in vitro and divided into the following groups:cell control group,γ-BGT group,carbachol group,gallamine group,γ-BGT+H-89 group,and γ-BGT+Y-27632 group.The γ-BGT group,carbachol group,and gallamine group were incubated with γ-BGT(40 mg·L-1),carbachol(100 mmol·L-1),and gallamine(100 mmol·L-1),respectively,for 4 h.The γ-BGT+H-89 and γ-BGT+Y-27632 groups were pretreated with γ-BGT(40 mg·L-1)for 4 h,followed by incubation with the protein kinase A(PKA)inhibitor H-89(50 mmol·L-1)and the Ca2+channel inhibitor Y-27632(50 mmol·L-1)for another 2 h,respectively.ELISA was used to measure the levels of Glu,GABA,cAMP,and calpain in the primary mouse medul-lary neurons.Western blotting was employed to assess the protein expression levels of GAD65 and GAD67,and PKA phosphorylation levels.Fluo-4 fluorescent probe was used to detect the intracellular Ca2+level.RESULTS The respiratory rate of mice significantly decreased after iv administration of γ-BGT(γ-BGT group)(P<0.05).After treatment with nikethamide(nikethamide group),the respiratory rate significantly recovered(P<0.05).Compared with the normal control group,the γ-BGT group exhib-ited a significant decrease in Glu content(P<0.05),a significant increase in GABA content(P<0.05),and a significant decrease in the Glu/GABA ratio.Additionally,the protein expression levels of GAD65 and GAD67 were significantly elevated(P<0.05).Compared with the γ-BGT group,the γ-BGT+niketh-amide group showed a significant increase in Glu content(P<0.05),a significant decrease in GABA content(P<0.05),a significant increase in the Glu/GABA ratio,and a significant reduction in GAD65 and GAD67 protein expression levels(P<0.05).Compared to the cell control group,the γ-BGT group demonstrated a significant decrease in Glu content(P<0.05),a significant increase in GABA content(P<0.05),and a significant reduction in the Glu/GABA ratio.Furthermore,the protein expression levels of GAD65 and GAD67 were significantly elevated(P<0.05).Additionally,cAMP content,PKA phosphor-ylation levels,Ca2+levels,and calpain activity were all significantly increased(all P<0.05).Glu,GABA,Glu/GABA ratio,and GAD expression levels in the γ-BGT group changed in the same way as in the gallamine group;In the γ-BGT+Y-27632 group,calpain activity and expression levels of GAD65 and GAD67 were all significantly decreased(all P<0.05).In the γ-BGT+H-89 group,Ca2+levels and calpain activity were significantly reduced(all P<0.05).CONCLUSION γ-BGT-induced poisoning can lead to respiratory distress in mice,possibly through the antagonism of M2 muscarinic acetylcholine receptors in medullary neurons,activation of the cAMP/PKA signaling pathway,elevation of intracellular Ca2+levels,and increased expression and activity of GAD,resulting in an imbalance of Glu and GABA in the medulla.

Hypercholesterolemia is a significant risk factor for the development of atherosclerosis. 2',3',5'-Tri-O-acetyl-N ⁶-(3-hydroxyphenyl) adenosine (IMM-H007), a novel AMPK agonist, has shown protective effects in metabolic diseases. However, its impact on cholesterol and triglyceride metabolism in hypercholesterolemia remains unclear. In this study, we aimed to elucidate the effects and specific mechanisms by which IMM-H007 regulates cholesterol and triglyceride metabolism. To achieve this goal, we used Apoe ^-/- and Ldlr ^-/- mice to establish a hypercholesterolemia/atherosclerosis model. Additionally, hepatocyte-specific Ampka1/2 knockout mice were subjected to a 5-week high-cholesterol diet to establish hypercholesterolemia, while atherosclerosis was induced via AAV-PCSK9 injection combined with a 16-week high-cholesterol diet. Our results demonstrated that IMM-H007 improved cholesterol and triglyceride metabolism in mice with hypercholesterolemia. Mechanistically, IMM-H007 modulated the AMPKα1/2-LDLR signaling pathway, increasing cholesterol uptake in the liver. Furthermore, IMM-H007 activated the AMPKα1-FXR pathway, promoting the conversion of hepatic cholesterol to bile acids. Additionally, IMM-H007 prevented hepatic steatosis by activating the AMPKα1/2-ATGL pathway. In conclusion, our study suggests that IMM-H007 is a promising therapeutic agent for improving hypercholesterolemia and atherosclerosis through the activation of AMPKα.