[摘 要] 目的：探讨DNA甲基转移酶1（DNMT1）通过稳定zeste基因增强子同源物2（EZH2）促进结直肠癌（CRC）HCT8细胞增殖与迁移的机制。方法：利用生物信息学方法分析DNMT1在CRC组织中的表达水平。WB法检测DNMT1在CRC细胞HCT8、SW620和正常结肠上皮细胞NCM460中的表达。通过siRNA或慢病毒载体转染HCT8细胞，分为siNC组、siDNMT1组、Vector组、DNMT1-OE组、siTRAF6组、siEZH2组、siEZH2 + DNMT1-OE组。采用克隆形成实验、CCK-8法、Transwell实验和划痕愈合实验检测敲低或过表达DNMT1对HCT8细胞增殖与迁移的影响，WB和qPCR法检测EZH2蛋白和mRNA水平，免疫沉淀（IP）法检测EZH2泛素化水平，免疫荧光双染检测肿瘤坏死因子受体相关因子6（TRAF6）与EZH2的细胞内共定位情况，克隆形成和划痕愈合实验验证EZH2对DNMT1功能的逆转作用。收集2022—2025年间郑州大学附属洛阳中心医院手术切除的12例CRC患者的癌及癌旁组织标本，采用免疫组化法检测CRC组织中DNMT1、TRAF6和EZH2的表达水平。结果：DNMT1在CRC组织中表达显著高于癌旁组织（P < 0.01），且在CRC细胞中表达上调（P < 0.05）；DNMT1敲低显著抑制HCT8细胞增殖及迁移（均P < 0.01），过表达则相反（均P < 0.01）。DNMT1正向调控EZH2的蛋白水平（P < 0.01），而mRNA水平不变（P > 0.05）。MG132可恢复siDNMT1组的EZH2蛋白表达（P < 0.01），且siDNMT1组EZH2泛素化水平升高。DNMT1负向调控TRAF6的表达（P < 0.01），且TRAF6与EZH2在细胞质中共定位，IP证实两者直接结合。敲低TRAF6可减弱EZH2的泛素化水平，敲低EZH2可逆转DNMT1对HCT8细胞增殖、迁移的促进作用（均P < 0.01）。DNMT1和EZH2在CRC组织中呈高表达（P < 0.01），TRAF6在CRC组织中表达显著低于癌旁组织（P < 0.05）。结论：DNMT1通过抑制TRAF6稳定EZH2促进CRC细胞的增殖和迁移，DNMT1、TRAF6和EZH2可能是CRC治疗的潜在靶点。

ObjectiveTo establish a model of dry eye with lung Yin deficiency syndrome in mice. MethodsA total of 40 SPF C57BL/6J mice were assigned via the random number table method into 5 groups （n=8）： Normal control， model control， and high-， medium-， and low-dose （11.7， 5.85， and 2.925 g·kg^-1， respectively） Yangyin Qingfeitang. Mice in the normal control group were fed normally without any intervention. Mice in Yangyin Qingfeitang group and model control group were treated with 0.2% benzalkonium chloride eye drops （5 μL） twice a day and fed in a controlled drying system in a dry environment for 28 days. At the same time， the mice were administrated with thyroxine tablet solution by gavage and placed in a glass fumigation tank （SO₂ concentration： 0.5 g·m^-3） for 14 days. After 4 weeks， mice in Yangyin Qingfeitang groups were treated with Yangyin Qingfeitang by gavage and those in the normal control group and model control group were administrated with deionized water at 0.01 mL·g^-1. The body mass， anal temperature， four examination information （claw and nail appearance）， basic tear secretion test， tear film rupture time， corneal fluorescein staining， and lacrimal gland HE staining were compared among groups. Compound Yangyin Qingfeitang granules were used to measure the syndrome to verify the success of modeling. ResultsAfter 28 days of continuous modeling， compared with the normal control group， the model group exhibited listless and emaciated status， coughing， drowsiness， dry and dull hair， dry and hard stool， reduced food intake and water intake， red lip circumference， red tongue with reduced fluid， dry nose and teeth， red claws and nails， body mass gain， decreased anal mild tear secretion （P<0.05）， and shortened tear film rupture time （P<0.05）. After 28 days of modeling， the mice showed large corneal fluorescein staining range， severe corneal injury， and increased content of interleukin （IL）-18， IL-β， and tumor necrosis factor （TNF）-α in lacrimal gland， compared with those in the normal control group （P<0.05）. After the treatment with Yangyin Qingfeitang， the mice had good drinking and eating conditions， with lighter redness of the tongue， moist nose， moist and shiny teeth， and the claw and nail color close to that in the normal group. Compared with the model control group， Yangyin Qingfeitang groups showed increases in body mass and anal temperature （P<0.05）， tear secretion （P<0.05）， and tear film rupture time （P<0.05）， narrowed range of corneal fluorescein staining， and declined levels of IL-18， IL-β， and TNF-α in lacrimal glands （P<0.01）. The high-dose group had the best effect， with the indicators close to the levels in the normal control group. ConclusionThe animal model of dry eye with lung Yin deficiency syndrome can be established by culture in a controlled drying system， treatment with benzalkonium chloride eye drops for 28 days， and administration of thyroxine tablet solution combined with SO₂ fumigation for 14 days.

ObjectiveTo investigate the mechanisms of Runmu Dihuang decoction （RMDHD） in treating perimenopausal dry eye with liver-kidney Yin deficiency syndrome based on the silent information regulator 3 （SIRT3）/hypoxia-inducible factor-1α （HIF-1α）/nuclear factor-κB （NF-κB） signaling pathway. MethodsSixty female Sprague-Dawley rats were randomly divided into six groups （n=10 per group）： Sham operation group， model group， sodium hyaluronate eye drop group， and low-， medium-， and high-dose RMDHD groups （5.625， 11.25， 22.50 g·kg^-1）. Except for the sham operation group， all rats underwent bilateral ovariectomy and were administered 0.1% benzalkonium chloride eye drops combined with long-term chronic irritation to establish a perimenopausal dry eye model with liver-kidney Yin deficiency syndrome. Drug administration began in the 11th week after modeling and continued for 21 days. General conditions， screen-grip test scores， tear secretion volume， tear film breakup time （TFBUT）， and corneal fluorescein staining were recorded. Serum levels of reactive oxygen species （ROS）， follicle-stimulating hormone （FSH）， estradiol （E₂）， and progesterone （PROG） were measured by enzyme-linked immunosorbent assay （ELISA）. Pathological changes in the lacrimal glands， corneas， and uteri were observed using hematoxylin-eosin （HE） staining. Protein expression levels of SIRT3， HIF-1α， phosphorylated NF-κB p65 （p-NF-κB p65）， and total NF-κB p65 in the lacrimal glands were detected by Western blot. The expression of inflammatory cytokines interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in the lacrimal glands was assessed by immunohistochemistry （IHC）. ResultsAfter model establishment， no significant differences were observed among the groups except the sham operation group. Compared with the sham operation group， the other groups exhibited slowed movement， dull responses， increased irritability， reduced body weight， elevated rectal temperature， decreased screen-grip test scores， reduced tear secretion， and significantly shortened TFBUT （P<0.05）. After treatment， compared with the model group， the sodium hyaluronate eye drop group and all RMDHD groups showed improved general conditions， significantly increased tear secretion （P<0.05）， prolonged TFBUT （P<0.05）， and elevated screen-grip test scores （P<0.05）. Serum ROS and FSH levels were significantly decreased， while E₂ and PROG levels were significantly increased （P<0.05）. Pathological damage to the cornea， lacrimal glands， and uterus was ameliorated. In addition， protein expression levels of SIRT3 and HIF-1α in the lacrimal glands were significantly upregulated （P<0.05）， whereas the expression of p-NF-κB p65， IL-1β， and TNF-α was significantly downregulated （P<0.05）. ConclusionRMDHD increases tear secretion and TFBUT， improves lacrimal gland and corneal injury， and alleviates dry eye symptoms in a perimenopausal dry eye rat model with liver-kidney Yin deficiency syndrome. The underlying mechanism may be related to regulation of the SIRT3/HIF-1α/NF-κB signaling pathway， inhibition of oxidative stress and inflammatory responses， and reduction of ocular surface tissue damage.

BackgroundChronic insomnia disorder has become a significant public health issue， and it may be associated with dyslipidemia. Previous studies on dyslipidemia in patients with chronic insomnia disorder have mainly focused on exploring the relationship between subjective short sleep duration and dyslipidemia， while there have been limited studies on the relationship between objective short sleep duration and dyslipidemia. ObjectiveTo explore the relationship between objective short sleep duration and dyslipidemia in patients with chronic insomnia disorder， in order to provide references for the prevention and intervention of dyslipidemia in this population. MethodsA total of 103 patients who were hospitalized at The Third Hospital of Mianyang from August 2022 to November 2023 and met the diagnostic criteria for chronic insomnia disorder as defined in the International Classification of Sleep Disorder， third edition （ICSD-3） were retrospectively collected. The objective sleep duration of the patients was obtained through polysomnography. The patients were divided into two groups based on their objective sleep duration： the group with objective sleep duration ≥ 7 hours （n=71） and the group with objective sleep duration < 7 hours （n=32）. Binary Logistic regression analysis was used to explore the impact of objective sleep duration < 7 hours on dyslipidemia. ResultsAmong 103 patients with chronic insomnia disorder， 59 cases （57.28%） were identified with dyslipidemia. The comparison of dyslipidemia conditions between the group with objective sleep duration ≥ 7 hours and the group with objective sleep duration < 7 hours showed a statistically significant difference （χ²=5.956， P<0.05）. Compared with the group with objective sleep duration ≥7 hours， the group with objective sleep duration < 7 hours exhibited significantly lower high-density lipoprotein cholesterol levels， and reduced sleep efficiency （t=-2.003， -5.482， P<0.05 or 0.01）. Binary Logistic regression analysis results showed that the risk of abnormal blood lipids in patients with chronic insomnia disorder with objective sleep duration < 7 hours was 3.128 times higher than that of patients with objective sleep duration ≥ 7 hours （OR=3.128， 95% CI： 1.139–8.588）. ConclusionObjective short sleep duration may be a risk factor for dyslipidemia in patients with chronic insomnia disorder.

BackgroundAnxiety symptoms have become a public health issue affecting the physical and mental health of the elderly population. Mental health literacy is a predictor of anxiety symptoms in the elderly. Currently， there are limited studies on the pathogenic mechanism between the two. Exploring the relationship and mechanism between mental health literacy and anxiety symptoms among the elderly is of great significance for improving the mental health level of the elderly. ObjectiveTo investigate the impact of mental health literacy on anxiety symptoms in the elderly， and to analyze the role of insomnia in this process， in order to provide references for the formulation of prevention and intervention strategies for anxiety symptoms in the elderly. MethodsFrom August 2021 to December 2022， a total of 10 650 older adults aged 60 years old and above were selected from a city in Sichuan Province using a multistage stratified sampling method. Participants completed the self-compiled demographic questionnaire， the Insomnia Severity Index （ISI）， the Generalized Anxiety Disorder Scale-7 item （GAD-7）， and the National Mental Health Literacy Questionnaire （NMHLQ）. Spearman correlation analysis was adopted to examine the correlation between the scores of the scales. Model 4 in SPSS 27.0 plugin Process 4.1 was employed to test the pathway of insomnia between mental health literacy （and its various dimensions） and anxiety symptoms. ResultsAmong the participants， 9 609 cases （90.23%） completed the valid questionnaire survey， and 1 680 cases （17.48%） were found to have anxiety symptoms. The total score of the NMHLQ for the elderly， as well as the scores of the knowledge， awareness， and skills dimensions， were negatively correlated with the GAD-7 score and the ISI score （r_s=-0.506–-0.054， P<0.01）， and the ISI score was positively correlated with the GAD-7 score （r_s=0.666， P<0.01）. Insomnia served as the mediating pathway between the mental health literacy and anxiety symptoms， with an indirect effect value of -0.210 （95% CI： -0.227–-0.193）， accounting for 54.97% of the total effect. Insomnia was the mediating pathway between the mental health literacy knowledge and anxiety symptoms of the elderly， with an indirect effect value of -0.161 （95% CI： -0.178–-0.144）， accounting for 52.61% of the total effect. Insomnia played a mediating role in the relationship between the awareness of mental health literacy and anxiety symptoms， with an indirect effect value of -0.323 （95% CI： -0.342–-0.302）， accounting for 76.36% of the total effect. Insomnia was the mediating pathway between the mental health literacy skills and anxiety symptoms of the elderly， with an indirect effect value of -0.172 （95% CI： -0.187–-0.159）， accounting for 53.75% of the total effect. ConclusionThe dimensions of mental health literacy， knowledge， awareness and skills of the elderly not only directly affect anxiety symptoms， but also indirectly influence anxiety symptoms through the pathway of insomnia.［Funded by Medical Research Project Plan of Sichuan Province （number， S23049）］

ObjectiveTo explore the diagnostic value of resting-state functional magnetic resonance imaging (rs-fMRI) for mild cognitive impairment (MCI) stage of Alzheimer's disease (AD), and to investigate the changes in brain functional connectivity in default mode network (DMN) and medial temporal lobe (MTL) regions. MethodsLiteratures were retrieved from multiple databases including CNKI, PubMed, Medline, and Embase, etc. from January, 2020 to October, 2025, investigating the changes in functional connectivity of DMN and MTL in patients with MCI compared to health control (HC) using rs-fMRI. Two researchers independently screened and extracted the literatures and methodological quality was assessed using QUADAS-2. Review Manager 5.4 was used to perform a meta-analysis of neuroimaging indicators in MCI patients and HC subjects. Stata and SDM were utilized to summarize diagnostic efficacy and brain functional connectivity, calculating the over all sensitivity, specificity and summary receiver operating characteristic-area under the curve (SROC-AUC). Deeks funnel plots were drawn, literature weights were analyzed, and subgroup analyses were conducted. ResultsA total of ten literatures were ultimately included, involving 1 010 patients with MCI and 1 714 HC subjects. MCI patients showed decreased or abnormal blood oxygen level dependence signals in DMN, focusing on bilateral medial prefrontal cortex (mPFC) and posterior cingulate cortex. The SROC-AUC was 0.91. In MTL, there was a characteristic decrease of spontaneous neural functional connectivities between the hippocampus and other regions, reflecting the obstruction of information transmission from episodic memory to the central nodes. Abnormal functional connectivity in DMN and MTL resulted in compensatory resting-state functional connectivity enhancement in other subnetworks or local functional connections, such as frontoparietal network and the hippocampal-parietal network. Abnormal activation of mPFC suggested atrophy of the hippocampus or abnormal brain function. The decline in functional connectivity between DMN and MTL indicated impairment of memory and information processing in the early stage. ConclusionThe early functional decoupling between DMN and MTL is a crucial neural mechanism at the MCI stage of AD, providing important neuroimaging evidence for the early diagnosis of AD.

ObjectiveTo quantitatively evaluate the effect of phrenic nerve-abdominal muscle electrical stimulation on pulmonary ventilation distribution in stroke patients using electrical impedance tomography (EIT). MethodsThirty-five stroke patients were enrolled at the First Affiliated Hospital of Chongqing Medical University from September, 2024 to June, 2025, and all received standardized phrenic nerve-abdominal muscle electrical stimulation. Percentage of ventilation in gravity-dependent regions of interesting (ROI%), center of ventilation (COV), global inhomogeneity index (GI) and change in end-expiratory lung impedance (ΔEELI) were measured with EIT monitoring before treatment (T0), immediately after treatment (T1), and at five, ten, 15, 20, 25, 30, 40 and 50 minutes after treatment (T2 to T9). ResultsThere were significant differences in ROI% (F = 7.003, P < 0.001) and COV (F = 5.722, P < 0.001) at different time points, both peaking at T1, followed by a downward trend until T5. No significant differences were observed in GI (F = 1.849, P = 0.097) and ΔEELI (F = 0.208, P = 0.871) across time points; however, GI at T7 was lower than that at T0 (P < 0.05). ConclusionPhrenic nerve-abdominal muscle electrical stimulation can improve the ventilation ratio in gravity-dependent regions and shift the center of ventilation dorsally. The improvement in ventilation distribution generally peaks at the end of treatment and lasts for approximately 20 minutes.

ObjectiveUlcerative colitis is a prevalent immunoinflammatory disease. Th17/Treg cell imbalance and gut microbiota dysregulation are key factors in ulcerative colitis pathogenesis. The actin cytoskeleton contributes to regulating the proliferation, differentiation, and migration of Th17 and Treg cells. Wdr63, a gene containing the WD repeat domain, participates in the structure and functional modulation of actin cytoskeleton. Recent research indicates that WDR63 may serve as a regulator of cell migration and metastasis via actin polymerization inhibition. This article aims to explore the effect of Wdr63 deletion on Th17/Treg cells and ulcerative colitis. MethodsWe constructed Wdr63^-/- mice, induced colitis in mice using dextran sulfate sodium salt, collected colon tissue for histopathological staining, collected mesenteric lymph nodes for flow cytometry analysis, and collected healthy mouse feces for microbial diversity detection. ResultsCompared with wild-type colitis mice, Wdr63^-/- colitis mice had a more pronounced shortening of colonic tissue, higher scores on disease activity index and histological damage index, Treg cells decreased and Th17 cells increased in colonic tissue and mesenteric lymph nodes, a lower level of anti-inflammatory cytokine IL-10, and a higher level of pro-inflammatory cytokine IL-17A. In addition, WDR63 has shown positive effects on maintaining intestinal microbiota homeostasis. It maintains the balance of Bacteroidota and Firmicutes, promoting the formation of beneficial intestinal bacteria linked to immune inflammation. ConclusionWdr63 deletion aggravates ulcerative colitis in mice, WDR63 inhibits colonic inflammation likely by regulating Th17/Treg balance and maintains intestinal microbiota homeostasis.

OBJECTIVE: To investigate the differences in indwelling duration, clinical scenarios, and complications between the modified midline catheter (MMC) and the central venous catheter (CVC) in the treatment of patients in the medical intensive care unit (ICU) and the risk factors for complications based on real-world data. METHODS: A retrospective cohort study was conducted. The adult patients admitted to the medical ICU of the Third Xiangya Hospital of Central South University and had undergone placement of either a MMC or a CVC between January 1, 2023, and July 31, 2024, were consecutively enrolled by querying the hospital's electronic medical record system. Based on the type of catheter inserted, the patients were divided into the MMC group and the CVC group. The two groups were compared regarding the selection of catheters in the context of different underlying diseases, the actual clinical application after catheterization, catheter-related complications, the international normalized ratio (INR) and platelet count (PLT) during puncture and catheterization, the length of ICU stay, total length of hospital stay, catheter indwelling duration, and mortality during hospitalization. Multivariate Logistic regression analysis was employed to identify independent risk factors for catheter removal. RESULTS: Among the 274 patients, 52 received a MMC and 222 received a CVC. The utilization rate of MMC was significantly higher than that of CVC in patients with acute respiratory distress syndrome (ARDS), cardiovascular disease, and cancer [ARDS: 92.3% (48/52) vs. 70.3% (156/222), cardiovascular disease: 84.6% (44/52) vs. 54.5% (121/222), cancer: 30.8% (16/52) vs. 17.1% (38/222), all P < 0.05]. However, the use of MMC was significantly lower than CVC when vasoactive drug infusion was required [57.7% (30/52) vs. 79.7% (177/222), P < 0.05]. A significantly higher proportion of patients in the MMC group had a catheter indwelling time ≥ 12 days as compared with the CVC group [32.7% (17/52) vs. 13.5% (30/222), P < 0.05]. There were no statistically significant differences in other underlying diseases, venous access usage, INR and PLT during puncture and catheterization, length of ICU stay, total length of hospital stay, and in-hospital mortality of patients between the two groups. Regarding catheter-related complications, although the incidence of partial or complete catheter removal in the MMC group was significantly higher than that in the CVC group [36.5% (19/52) vs. 5.4% (12/222), P < 0.05], the incidence of puncture site fluid leakage, puncture site skin allergy, and deep vein thrombosis were significantly lower than those in the CVC group [puncture site fluid leakage: 1.9% (1/52) vs. 22.1% (49/222), puncture site skin allergy: 0% (0/52) vs. 20.7% (46/222), deep vein thrombosis: 3.8% (2/52) vs. 16.7% (37/222), all P < 0.05]. Furthermore, the proportion of patients experiencing three or more types of complications in the MMC group was significantly lower than that in the CVC group [5.8% (3/52) vs. 17.6% (39/222), P < 0.05]. Multivariate Logistic regression analysis of risk factors for catheter removal identified the use of a MMC [odds ratio (OR) = 8.518, 95% confidence interval (95%CI) was 3.710-19.560, P < 0.001] and a catheter indwelling time ≥ 12 days (OR = 3.133, 95%CI was 1.297-7.567, P = 0.011) as independent risk factors. CONCLUSIONS MMC was more frequently used in patients with ARDS, cardiovascular disease, and cancer, whereas CVC was primarily employed for vasoactive drug infusion. The use of MMC and a longer indwelling time were identified as independent risk factors for catheter removal. Despite a higher removal rate, the overall incidence of complications was significantly lower with MMC than with CVC. These findings suggest that MMC could serve as a routine alternative to CVC in most of clinical scenarios, provided that measures are implemented to prevent removal.
Humans ; Retrospective Studies ; Intensive Care Units ; Catheterization, Central Venous/methods* ; Central Venous Catheters ; Risk Factors ; Length of Stay ; Male ; Female ; Middle Aged ; Adult ; Catheters, Indwelling ; Aged