OBJECTIVE To investigate the mechanism of isochlorogenic acid A against hepatocellular carcinoma based on the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway and multi-omics technology. METHODS The invasion rate and migration rate of human hepatocellular carcinoma HepG2 cells after 48 h of intervention with 0 （control group）， 0.25 and 0.5 mg/mL isochlorogenic acid A were examined； mRNA expression of DEP domain-containing mTOR-interacting protein （DEPTOR）， the protein expressions of mTOR， PI3K and phosphatase and tensin homologue deleted on chromosome ten （PTEN）， as well as the phosphorylation level of Akt protein were determined in the cells. Metabolomics analysis was performed using liquid chromatography-tandem mass spectrometry， and differential metabolites were screened and subjected to Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis； transcriptomics monitoring was conducted by RNA sequencing， and differentially expressed genes were screened and subjected to gene ontology （GO） and KEGG pathway enrichment analyses. RESULTS Compared with the control group， intervention with 0.25 and 0.5 mg/mL isochlorogenic acid A for 48 h significantly inhibited the invasion rate and migration rate of HepG2 cells， significantly up-regulated the mRNA expression of DEPTOR and the protein expression of PTEN， and significantly down-regulated the protein expression of PI3K and the phosphorylation level of Akt protein （except for 0.25 mg/mL isochlorogenic acid A） （ P ＜0.05）. A total of 304 differential metabolites and 212 differentially expressed genes were screened by multi-omics analysis. KEGG pathway enrichment analysis suggested that isochlorogenic acid A regulated key signaling of HepG2 cell growth mainly by inhibiting the PI3K/Akt signaling pathway， synergizing with metabolic reprogramming such as mTOR signaling pathway， ferroptosis， pentose phosphate pathway and purine/pyrimidine metabo lism. CONCLUSIONS The anti-hepatocellular carcinoma effect of isochlorogenic acid A is associated with the blockade of abnormal activation of the PI3K/Akt/mTOR signaling pathway. In addition， it may also be related to the inhibition of the pentose phosphate pathway and purine/pyrimidine metabolism， as well as the induction of ferroptosis，etc.

OBJECTIVES: To investigate the effect of BRD4 inhibition on migration of esophageal squamous cell carcinoma (ESCC) cells with low acetyl-CoA carboxylase 1 (ACC1) expression. METHODS: ESCC cell lines with lentivirus-mediated ACC1 knockdown or transfected with a negative control sequence (shNC) were treated with DMSO, JQ1 (a BRD4 inhibitor), co-transfection with shNC-siBRD4 or siNC with additional DMSO or C646 (an ahistone acetyltransferase inhibitor) treatment, or JQ1combined with 3-MA (an autophagy inhibitor). BRD4 mRNA expression in the cells was detected using RT-qPCR. The changes in cell proliferation, migration, autophagy, and epithelial-mesenchymal transition (EMT) were examined with CCK8 assay, Transwell migration assay, and Western blotting. RESULTS: ACC1 knockdown did not significantly affect BRD4 expression in the cells but obviously increased their sensitivity to JQ1. JQ1 treatment at 1 and 2 μmol/L significantly inhibited ESCC cell proliferation, while JQ1 at 0.2 and 2 μmol/L promoted cell migration. The cells with ACC1 knockdown and JQ1 treatment showed increased expresisons of vimentin and Slug and decreased expression of E-cadherin. BRD4 knockdown promoted migration of ESCC cells, and co-transfection with shACC1 and siBRD4 resulted in increased vimentin and Slug expressions and decreased E-cadherin expression in the cells. C646 treatment of the co-transfected cells reduced acetylation levels, decreased vimentin and Slug expressions, and increased E-cadherin expression. Treatment with JQ1 alone obviously increased LC3A/B-II levels in the cells either with or without ACC1 knockdown. In the cells with ACC1 knockdown and JQ1 treatment, additional 3-MA treatment significantly decreased the expressions of vimentin, Slug and LC3A/B-II and increased the expression of E-cadherin. CONCLUSIONS BRD4 inhibition promotes autophagy of ESCC cells via a histone acetylation-dependent mechanism, thereby enhancing EMT and ultimately increasing cell migration driven by ACC1 deficiency.
Humans ; Cell Movement ; Transcription Factors/metabolism* ; Esophageal Neoplasms/metabolism* ; Cell Line, Tumor ; Cell Cycle Proteins ; Azepines/pharmacology* ; Epithelial-Mesenchymal Transition ; Carcinoma, Squamous Cell/metabolism* ; Esophageal Squamous Cell Carcinoma ; Triazoles/pharmacology* ; Nuclear Proteins/genetics* ; Cell Proliferation ; Acetyl-CoA Carboxylase/genetics* ; Transfection ; Autophagy ; Bromodomain Containing Proteins

Lentiviral vectors(LVs)are key gene delivery tools for integrating target genes into the host genome,but they may also pose risks of insertional mutagenesis.The vector copy number(VCN)in cells is critical for determining the safety of gene modification.However,the reliability and accuracy of its quantification process are influenced by multiple factors.Developing cell reference materials with specific vector copy numbers represents a viable approach to enhance the reliability and consistency of measurement results,enabling quality control of the quantification process and traceability of outcomes.However,the preparation of such reference materials faces challenges in cell sample design,preparation protocols,and advanced quantification techniques.In this study,T lymphocyte cell line Jurkat-based reference materials with LV gene copy numbers of 1 and 2 copy/cell were developed.A high-precision duplex digital polymerase chain reaction(dPCR)method was established to quantify the LV gene and endogenous genes simultaneously.Additionally,the results of dPCR were cross-validated through next-generation sequencing and flow cytometric analysis.Ultimately,confocal microscopy characterization results showed that the developed cell reference materials had intact morphology.The quantification result of VCN-1 was(1.07±0.11)copy/cell,and that of VCN-2 was(2.09±0.21)copy/cell.These cell reference materials demonstrated compliance with stability and homogeneity requirements,and could be applied for quality control throughout the VCN measurement workflow and metrological traceability,improving the accuracy,comparability,and validity of copy number measurements.

The chemical components of Carthami Flos were investigated by using macroporous resin, silica gel column chromatography, reversed-phase octadecylsilane(ODS) column chromatography, Sephadex LH-20, and semi-preparative high-performance liquid chromatography(HPLC). The planar structures of the compounds were established based on their physicochemical properties and ultraviolet-visible(UV-Vis), infrared(IR), high-resolution electrospray ionization mass spectrometry(HR-ESI-MS), and nuclear magnetic resonance(NMR) spectroscopic technology. The absolute configurations were determined by comparing the calculated and experimental electronic circular dichroism(ECD). Six flavonoid C-glycosides were isolated from the 30% ethanol elution fraction of macroporous resin obtained from the 95% ethanol extract of Carthami Flos, and identified as saffloquinoside F(1), 5-hydroxysaffloneoside(2), iso-5-hydroxysaffloneoside(3), isosafflomin C(4), safflomin C(5), and vicenin 2(6). Among these, the compounds 1 to 3 were new chalcone C-glycosides. The compounds 1, 2, 4, and 5 could significantly increase the viability of H9c2 cardiomyocytes damaged by oxygen-glucose deprivation/reoxygenation(OGD/R) at a concentration of 50 μmol·L~(-1), showing their good cardioprotective activity.
Glycosides/pharmacology* ; Flowers/chemistry* ; Drugs, Chinese Herbal/pharmacology* ; Carthamus tinctorius/chemistry* ; Chalcones/pharmacology* ; Animals

Objective To investigate the status and influencing factors of type 2 diabetes mellitus(T2DM)patients' fear of complications,and to provide a reference for formulating targeted intervention measures.Methods From April to November 2024,370 patients with T2DM in 2 tertiary general hospitals in Daqing City were selected by convenience sampling method.General data questionnaire,Fear of Complications Questionnaire,Self-Perceived Burden Scale,Psychological Capital Questionnaire,Mishel Uncertainty in Illness Scale and Family Apgar Index Questionnaire were used for investigation.Univariate analysis and binary Logistic regression were performed to analyze the influencing factors.Results A total of 364 valid questionnaires were collected,with an effective recovery rate of 98.38％.The score of Fear of Complications Questionnaire was(23.47±7.47),and the incidence of fear of complications was 22.25％.Logistic regression analysis showed that medical payment methods,the number of complications,positive psychological capital and family care were the influencing factors of FoC in T2DM patients.Conclusion The fear of complications in T2DM patients is at a moderate level.Nursing staff should pay attention to the early assessment of patients' fear of complications,promptly identify and take effective measures to reduce the level of patients' fear of complications,improve their quality of life.

Objective:To analyze the incidence, clinical characteristics, and prognosis of basilar artery occlusion (BAO) patients presenting with paroxysmal sympathetic hyperactivity (PSH) as an initial symptom.Methods:A retrospective analysis was conducted on BAO patients with PSH manifestations who received endovascular treatment at Xingtai Central Hospital between January 2018 and August 2024. Demographic characteristics, clinical data, imaging findings, and follow-up information were collected.Results:A total of 136 BAO patients were included, of whom 22 cases (16.2%) were diagnosed with PSH. The age of the patients was (56.81±9.92) years, with males accounting for 86.4%(19/22). On admission, the Glasgow Coma Scale score was 4(3, 5), and the National Institutes of Health Stroke Scale score was 30(25, 35). The successful reperfusion (modified Thrombolysis In Cerebral Infarction 2b-3) rate was 90.9%(20/22). In-hospital mortality was 50.0%(11/22), symptomatic hemorrhagic transformation occurred in 4.5% (1/22), and any type of intracranial hemorrhage occurred in 9.1%(2/22). Clinical features of PSH included tachycardia, tachypnea, hypertension, fever, diaphoresis, and increased muscle tone. Imaging analysis showed a high proportion of cerebellar (100.0%, 22/22) and midbrain (72.7%, 16/22) involvement. The 90-day follow-up showed that 72.7%(16/22) of the patients had poor functional outcomes (modified Rankin Scale score of 4-6), and the 90-day mortality rate was 59.1%(13/22).Conclusions:Among patients with BAO undergoing endovascular therapy, the incidence of PSH as the initial manifestation was 16.2%(22/136). These patients were predominantly middle-aged men and commonly presented with tachycardia, tachypnea, hypertension, fever, diaphoresis, and increased muscle tone. Imaging findings mainly involved the cerebellum and midbrain. Despite a relatively high rate of recanalization, patients with PSH exhibited a higher risk of mortality and poor functional outcomes.

There is controversy over the specific and non-specific components in the efficacy evaluation of acupuncture therapy.The placebo effect,as a key non-specific factor,needs to be clarified in terms of its mechanism and clinical value.This article reviewed the research progress in the placebo effect of acupuncture from three aspects:cognitive process,influencing factors,and mechanism.It focused on the regulatory effects of patient expectations,doctor-patient interaction,and individual differences on the effect,summarized key mechanisms such as endogenous opioid system,dopamine reward pathway,and neural projection of rostral anterior cingulate cortex to pontine nucleus,and pointed out the methodological limitations of current comfort acupuncture designs,in order to provide theoretical basis for the optimization of acupuncture clinical research models and the deepening of the effect mechanism.

Objective:To investigate the influencing factors of adverse reactions of reconstructive flaps after postoperative precision radiation therapy for head and neck tumors.Methods:Medical records of 46 patients who underwent radiotherapy after flap reconstruction for head and neck tumors in the Affiliated Hospital of Inner Mongolia Medical University between January 2016 and October 2022 were retrospectively analyzed. The overall survival (OS), local regional control (LRC) and dosimetric parameters after radiotherapy were analyzed. The adverse reactions mainly including radiation dermatitis and flap necrosis at 3 and 6 months after radiotherapy, flap atrophy, flap fibrosis, dysphagia and chewing dysfunction at 12 and 24 months after radiotherapy were recorded. Adverse reactions were graded using the common terminology criteria for adverse events (CTCAE) version 5.0. Quantitative data were analyzed by one-way analysis of variance or rank-sum test. Qualitative data were analyzed by chi-square test or Fisher's exact test. Multivariate analysis of influencing factors of adverse reactions was performed using binary logistic stepwise regression.Results:All 46 patients were aged 57.6 years on average. The median follow-up time was 65 months (12-100 months). After the follow-up, 22 patients died, 6 recurred, and 7 had distant metastases. The 5-year OS rate was 48% and 5-year LRC rate was 69%. The incidence of radiation dermatitis was decreased over time after the end of radiotherapy: 80% (37/46) and 65% (30/46) at 3 months and 6 months after radiotherapy, respectively. The incidence of grade 3 radiation dermatitis was 11% (5/46) and 0 at 3 and 6 months after radiotherapy, respectively. The incidence of grade 2 flap necrosis was 4% (2/46) at 3 months after radiotherapy. At 12 months after radiotherapy, the incidence of flap atrophy and fibrosis was 83% (38/46) and 67% (31/46) , and the incidence of grade 3 flap atrophy and fibrosis was equally 4% (2/46) , respectively. All patients had dysphagia and chewing dysfunction at 12 months after radiotherapy. At 24 months after radiotherapy, 83% (38/46) and 61% (28/46) of patients still had dysphagia and chewing dysfunction. Multivariate regression analysis showed that irradiated flap volume, total radiotherapy dose, pharyngeal constrictor D mean and pharyngeal constrictor V 60 Gy, the interval between surgery and radiotherapy, gender and age were the independent influencing factors of adverse reactions of reconstructive flaps after precision radiation therapy for head and neck tumors. Conclusions:For patients undergoing flap reconstruction for head and neck tumors, under the premise of ensuring tumor control, the dose to normal tissue and flap blood supply area should be strictly limited, the irradiated volume should be reduced, and the interval between surgery and radiotherapy should be shortened (≤6 weeks) to reduce the risk of adverse reactions. Hypo-fractionated radiotherapy plan should be formulated or the total radiotherapy dose should be appropriately adjusted for elderly patients to reduce long-term complications.