Objective To establish a central cutting two-dimensional liquid chromatography for the determination of lamotrigine in human plasma. Methods External standard method was used. The first dimensional chromatographic column: SNCB（T）-1A（silica gel, 4.6 mm×50 mm, 5 μm）, mobile phase A:VCV-1D mobile phase, flow rate: 0.4 ml/min; mobile phase B: water, flow rate: 1.0 ml/min; second dimensional chromatographic column: Symmetry C₁₈ （4.6 mm×250 mm, 5 μm）, mobile phase: acetonitrile-10 mmol/L ammonium acetate solution（V/V=25∶75）, flow rate: 1.0 ml/min；Intermediate column: SBX 4-MA（resin, 3.0 mm×10 mm, 5µm）. The UV detection wavelength：306 nm, the column temperature：45 ℃, and the injection volume：200 μl. Results The linear range of lamotrigine was 1.24-39.50 μg/ml, the lower limit of quantification was 1.24 μg/ml, the detection limit was 0.02 μg/ml, the intra-day precision RSD was less than 5%, the day-to-day precision RSD was less than 10%, the variation of intra-day accuracy ranged from 102.17% to 111.17%, and the daytime accuracy variation ranged from 99.80% to 107.31% the recovery RSD was less than 5%, and the variation range was 89.95% -96.16%. After 24 hours storage at room temperature, repeated freezing and thawing for 3 times and storage at −40 ℃ for 2 weeks, the ratio of the measured value/labeled value ranged from 87.01% to 115.88%. Conclusion In this study, a method with simple operation, good stability, high sensitivity and good reproducibility was established, which could be suitable for clinical monitoring of blood concentration of lamotrigine and provide reliable monitoring data support for clinical individualized medication guidance.

Objective:To examine the therapeutic effects and molecular mechanisms of nuclear receptor NR4A1 in alleviating postoperative ileus (POI) in a rat model.Methods:Twenty-four Sprague-Dawley rats were randomly(random number) divided into four groups ( n=6/group): sham-operated control, POI model, POI model + NR4A1 stimultior (Cytosporone B, 13 mg/kg), and POI model + NR4A1 antagonist (DIM-C-pPhCO2Me, 2 mg/kg). After 24 hours, intestinal tissues and serum were collected for analysis. We assessed: (1) histopathological changes, (2) intestinal motility via propulsion rate, (3) NR4A1 expression by immunohistochemistry, (4) epithelial apoptosis via TUNEL assay, (5) inflammatory cytokines (IL-6, IL-4) by ELISA, (6) tight junction protein (occludin) by Western blot, and (7) p38MAPK/NF-κB pathway activation through combined western blot and immunofluorescence analyses. Results:Compared with sham controls, POI model rats showed (all P<0.05): significantly reduced NR4A1 expression, severe mucosal damage, increased inflammatory infiltration, elevated epithelial apoptosis, decreased occludin expression, impaired intestinal motility, upregulated pro-inflammatory cytokines (IL-6, IL-4), and activated p38MAPK/NF-κB signaling. NR4A1 activation with Cytosporone B significantly reversed these pathological changes (all P<0.05), while NR4A1 inhibition exacerbated them. Conclusions:NR4A1 activation attenuates POI by suppressing p38MAPK/NF-κB-mediated inflammation and preserving intestinal barrier function, suggesting its potential as a therapeutic target for postoperative ileus.

Objective To understand the clinical applicability and implementation of expert consensus on the implementation and removal of protective restraints in psychiatry,and to provide references for promoting the standardized practice of psychiatric protective restraints and updating the consensus.Methods By the convenience sampling method,a questionnaire survey was conducted among nurses from 480 hospitals in 30 provinces from June 15 to July 15,2024.The survey was conducted using the instrument for evaluating clinical applicability of guide-lines(version 2.0)and a self-compiled questionnaire on the clinical implementation of the restraint consensus.Results A total of 7,844 valid questionnaires were collected,with a valid questionnaire recovery rate of 93.78％.The results of clinical applicability scoring showed that the consensus had the lowest availability score(64.72％)and the highest acceptability score(76.74％).The results showed that nurses' receiving training and the level of their hospitals were the main influencing factors for scores in various dimensions(P＜0.05).4,774 participants(87.42％)believed that the application of consensus could enhance the standardization of nurses' restraint operations.The safety rate of the restraint consensus was 79.51％,and the economic ratio was 76.87％.Among the evaluators,1,739(22.17％)believed that there were implementation obstacles in the consensus.Conclusion The clinical applicability of the consensus is relatively good,and the application of the consensus helps to improve the standardization of clinical operations.In the future,efforts should be made to strengthen the promotion and training of the consensus,develop hierarchical promotion strategies according to the characteristics of medical institutions,and improve the quality of evidence for the consensus,so as to further enhance the clinical application effect of the consensus.

OBJECTIVE: To explore the reasons for false negative results by Optical genome mapping (OGM) analysis of three cases and propose strategies for handling them. METHODS: Three patients presented at the Affiliated Hospital of Nantong University between July 2022 and July 2024 were selected as study subjects. The patients included a 37-year-old female with two miscarriages, a 1.5-year-old boy with delayed motor development, and a 35-year-old male whose son had intellectual disability. The patients had undergone comprehensive evaluation with chromosomal karyotyping analysis, single nucleotide polymorphism microarray (SNP array) assay, fluorescence in situ hybridization (FISH), and methylation-specific multiple ligation-dependent probe amplification (MS-MLPA). A retrospective analysis was also carried out on the results of OGM testing. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.: 2020-K004). RESULTS: The chromosomal karyotype of patient 1 was 46,XX,4qs, and no abnormality was found by SNP array, FISH, and OGM testing. Patient 2 had a normal chromosomal karyotype, and SNP array analysis did not reveal any copy number abnormalities of chromosomal fragments but the presence of a homozygous region of approximately 79.58 Mb at 15q11.2-q26.3 (chr15: 22817871_102397317). MS-MLPA detection indicated that the copy number of the 15q11.2-q13 region was 2, and the degree of methylation was relatively high (average ratio = 1.0). OGM detection confirmed the presence of approximately 67.97 Mb of homozygosity in the chr15:33814680_101787650 [hg38] region of 15q14-q26.3. Patient 3 had a chromosomal karyotype of 46,XY,t(9;14)(q13;q11.2). No abnormality was found by OGM testing for patients 1 and 3. CONCLUSION As a novel cytogenetic technique, OGM can achieve high-resolution and high-precision analysis for numerical and structural genomic abnormalities. Nevertheless, it also has certain limitations, as its false negative results are related to factors such as the type of genomic variation, the chromosomal regions involved in the variation, the type of disease, and the version of human reference genome. Currently, it cannot be used as an independent method for the diagnosis of genetic diseases.
Humans ; Male ; Female ; Adult ; Polymorphism, Single Nucleotide/genetics* ; Karyotyping ; Chromosome Mapping/methods* ; Infant ; False Negative Reactions ; In Situ Hybridization, Fluorescence

Objective To investigate changes in the microRNA expression profiles of apoptotic vesicles(apoVs)derived from bone marrow mesenchymal stem cells(BMSCs)under a simulated negative pressure environment,and to provide a theoretical basis for understanding the mechanism by which mechanical stress microenvironments influence the progression of osteoarthritis.Methods A negative pressure cellular environment was established using a pressure-loading system.Cell viability and apoptosis were assessed via the CCK-8 assay,Western blotting,and Annexin V-FITC/PI double staining.ApoVs were isolated by differential centrifugation and characterized using transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Western blotting.Small RNA sequencing was performed using the HiSeq Single-End mode,and differential expression analysis of microRNAs was conducted using DESeq to screen for differentially expressed microRNAs.The differentially screened microRNAs were validated by real-time quantitative PCR.After treating BMSCs with inhibitors of these differentially expressed microRNAs,the effects of the screened microRNAs on BMSCs were detected.Results Compared to apoVs generated by BMSCs under STS chemical treatment,those produced under a-40 kPa pressure environment showed significantly upregulated miR-183-5p and downregulated miR-3473.GO and KEGG enrichment analyses revealed that these differentially expressed microRNAs affected cell activity and inflammatory responses through multiple signaling pathways.Inhibition of miR-183-5p and miR-3473 expression reduced the proliferative activity of BMSCs.After inhibiting miR-183-5p expression,the levels of inflammatory factors increased.Inhibition of miR-3473 expression did not alter the IL-6 expression level,but significantly increased the TNFα expression level.Conclusions MicroRNAs specifically expressed in BMSC-derived apoVs under negative pressure stimulation may act as critical mechanical signaling mediators,regulating inflammatory response processes to participate in the pathogenesis and progression of arthritis.

Objective To investigate changes in the microRNA expression profiles of apoptotic vesicles(apoVs)derived from bone marrow mesenchymal stem cells(BMSCs)under a simulated negative pressure environment,and to provide a theoretical basis for understanding the mechanism by which mechanical stress microenvironments influence the progression of osteoarthritis.Methods A negative pressure cellular environment was established using a pressure-loading system.Cell viability and apoptosis were assessed via the CCK-8 assay,Western blotting,and Annexin V-FITC/PI double staining.ApoVs were isolated by differential centrifugation and characterized using transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Western blotting.Small RNA sequencing was performed using the HiSeq Single-End mode,and differential expression analysis of microRNAs was conducted using DESeq to screen for differentially expressed microRNAs.The differentially screened microRNAs were validated by real-time quantitative PCR.After treating BMSCs with inhibitors of these differentially expressed microRNAs,the effects of the screened microRNAs on BMSCs were detected.Results Compared to apoVs generated by BMSCs under STS chemical treatment,those produced under a-40 kPa pressure environment showed significantly upregulated miR-183-5p and downregulated miR-3473.GO and KEGG enrichment analyses revealed that these differentially expressed microRNAs affected cell activity and inflammatory responses through multiple signaling pathways.Inhibition of miR-183-5p and miR-3473 expression reduced the proliferative activity of BMSCs.After inhibiting miR-183-5p expression,the levels of inflammatory factors increased.Inhibition of miR-3473 expression did not alter the IL-6 expression level,but significantly increased the TNFα expression level.Conclusions MicroRNAs specifically expressed in BMSC-derived apoVs under negative pressure stimulation may act as critical mechanical signaling mediators,regulating inflammatory response processes to participate in the pathogenesis and progression of arthritis.

Objective To understand the clinical applicability and implementation of expert consensus on the implementation and removal of protective restraints in psychiatry,and to provide references for promoting the standardized practice of psychiatric protective restraints and updating the consensus.Methods By the convenience sampling method,a questionnaire survey was conducted among nurses from 480 hospitals in 30 provinces from June 15 to July 15,2024.The survey was conducted using the instrument for evaluating clinical applicability of guide-lines(version 2.0)and a self-compiled questionnaire on the clinical implementation of the restraint consensus.Results A total of 7,844 valid questionnaires were collected,with a valid questionnaire recovery rate of 93.78％.The results of clinical applicability scoring showed that the consensus had the lowest availability score(64.72％)and the highest acceptability score(76.74％).The results showed that nurses' receiving training and the level of their hospitals were the main influencing factors for scores in various dimensions(P＜0.05).4,774 participants(87.42％)believed that the application of consensus could enhance the standardization of nurses' restraint operations.The safety rate of the restraint consensus was 79.51％,and the economic ratio was 76.87％.Among the evaluators,1,739(22.17％)believed that there were implementation obstacles in the consensus.Conclusion The clinical applicability of the consensus is relatively good,and the application of the consensus helps to improve the standardization of clinical operations.In the future,efforts should be made to strengthen the promotion and training of the consensus,develop hierarchical promotion strategies according to the characteristics of medical institutions,and improve the quality of evidence for the consensus,so as to further enhance the clinical application effect of the consensus.

Objective To evaluate the cost-utility of tiotropium/olodaterol in treating Chinese patients with moderate to very severe chronic obstructive pulmonary disease(COPD)and to provide references for selecting more economical inhaled preparations in clinical practice.Methods A four-state lifetime Markov model was established with a 3-month cycle.The health outcomes included life years and quality-adjusted life years.Costs,including direct medical costs,were calculated from the perspective of the Chinese health system.Discontinuation rates were derived from the discontinuation curve using GetData Graph Digitizer.The main output indicator of the model was the incremental cost-utility ratio,which was calculated from the queue simulation results to judge the economy of tiotropium bromide/odataterol.The scenario analysis and sensitivity analyses were carried out to detect the robustness of the base case result.Results Compared with tiotropium bromide,the patient treated with tiotropium bromide/odataterol gained an additional 0.0846 life years,an additional cost of ￥3 201.50,and additional 0.029 6 QALY.The incremental cost-utility ratio was 108 140.11 yuan/QALY,lower than the willingness-to-pay threshold of three times China's per capita GDP in 2021.The costs of tiotropium bromide and tiotropium bromide/odataterol had the greatest impact on the result in the one-way sensitivity analysis.93.8%of the Chinese COPD population was willing to pay for tiotropium bromide/odataterol under the threshold in the probability sensitivity analysis.Conclusion Tiotropium/olodaterol is a cost-effective alternative compared to tiotropium for patients with moderate to very severe COPD in China and the results were robust in the sensitivity analyses.

Objective To discuss the efficacy and safety of transarterial chemoembolization(TACE)and hepatic arterial infusion chemotherapy(HAIC)combined with tyrosine kinase inhibitors(TKI)and immune checkpoint inhibitors(ICI)for advanced hepatocellular carcinoma(HCC).Methods A total of 101 patients with unresectable HCC,who were admitted to the Affiliated Cancer Hospital of Harbin Medical University of China between January 2021 and October 2022 to receive treatment,were enrolled in this study.Of the 101 patients,50 received TACE+TKI+ICI therapy(TACE+TKI+ICI group)and 51 received HAIC+TKI+ICI therapy(HAIC+TKI+ICI group).The overall survival(OS)and the progression-free survival(PFS)were compared between the two groups,and the adverse events were analyzed to assess the safety of the therapeutic scheme.Results The median PFS in the TACE+TKI+ICI group was 12.0 months,which in the HAIC+TKI+ICI group was 11.0 months(P=0.030).The median OS was not achieved in the TACE+TKI+ICI group,which in the HAIC+TKI+ICI group was 14.6 months(P=0.005).The most common adverse effects in the TACE+TKI+ICI group were the elevation of total bilirubin(46.0％)and hepatic function injury(26.0％),which in the HAIC+TKI+ICI group were the decrease of albumin level(62.7％),fatigue(39.2％),and gastrointestinal reactions(31.4％).Conclusion For the treatment of advanced HCC,the therapeutic scheme of TACE+TKI+ICI has a better long-term survival benefits and the therapeutic scheme of HAIC+TKI+ICI can better maintain the liver function reserve of the patients.Neither therapeutic scheme shows any unexpected toxicity,and both therapeutic schemes have high clinical safety.(J Intervent Radiol,2024,33:543-548)

Gaucher Disease (GD) is an autosomal recessive lysosomal storage disorder characterized by high heterogeneity. This study aimed to further understand the correlation between clinical phenotypes and genotypes in GD patients through a retrospective analysis of 20 cases in Shanxi Bethune Hospital, including their clinical manifestations, laboratory tests, enzyme studies, and genetic results. Among the 20 GD patients, 16 were classified as Type Ⅰ GD with a median age of diagnosis of 24 years, and 4 were classified as Type Ⅲ GD with a median age of diagnosis of 19 years. All patients exhibited splenomegaly and thrombocytopenia, with 16 patients showing skeletal imaging changes, and 5 of them presenting with bone pain symptoms. Genetic analysis revealed 15 distinct mutations, predominantly missense mutations, with L483P being the most prevalent (35.7%), followed by V414L, L303I, and F252I. Mutation sites were predominantly located in exon 7. Noteworthy findings included the first report of the S310G mutation by our research group and the first occurrence of the K196R mutation in the Chinese population. Additionally, the N227S mutation was implicated in a potential association with neuropathy. Despite advancements, Uncertainties still exist in the correlation between clinical phenotypes and genotypes in GD patients.