Objective:To investigate the effects and potential mechanisms of melatonin on cognitive dysfunction induced by long-term anesthesia with isoflurane.Methods:Male C57BL/6J mice aged 2 months were divided into control group, isoflurane group, melatonin group, and isoflurane+ melatonin group by random number table, with 6 mice in each group.Three days after anesthesia, cognitive function of mice was assessed by Y-maze and fear conditioning (FC) tests. ATP content in the hippocampus was measured by an ATP assay kit. Western blot was used to detect the expression of DRP1, pDRP1, MFN2, pAMPK and SIRT1 proteins in the hippocampus. Cultured HT-22 cells derived from mouse hippocampal neurons in vitro were divided into control group, isoflurane group, melatonin group, and isoflurane + melatonin group, and the levels of reactive oxygen species (ROS) in each group were detected by flow cytometry after intervention. Statistical analysis was performed by SPSS 22.0 software, and one-way ANOVA was used for comparisons among multiple groups.Results:(1) There was a statistically significant difference in the percentage of freezing behavior in contextual fear memory among the four groups of mice ( F=39.09, P<0.05). The percentage of freezing behavior in the isoflurane group was lower than that in the control group ((44.23±8.88)% vs (75.87±5.90)%, P<0.05), while the percentage of freezing behavior in the isoflurane+ melatonin group((67.45±14.89)%)was higher than that in the isoflurane group ( P<0.05). There was also a statistically significant difference in the percentage of exploration in the novel arm among the four groups of mice ( F=13.87, P<0.05). The percentage of exploration in the novel arm in the isoflurane group was lower than that in the control group((33.64±6.53)% vs (47.13±3.87)%, P<0.05), while the percentage of exploration in the novel arm in the isoflurane+ melatonin group((43.05±1.64)%)was higher than that in the isoflurane group ( P<0.05). (2) Statistically significant difference in the levels of ATP in the hippocampus was found among the four groups of mice ( F=49.22, P<0.05). The level of ATP in the hippocampus in the isoflurane group was lower than that in the control group((2.29±0.15)nmol/mg vs (3.58±0.12)nmol/mg, P<0.05), while the level of ATP in the hippocampus in the isoflurane+ melatonin group ((3.02±0.27)nmol/mg)was higher than that in the isoflurane group ( P<0.05). There was a statistically significant difference in the levels of ROS in HT-22 cells among the four groups ( F=18.36, P<0.05). The level of ROS in HT-22 cells in the isoflurane group was higher than that in the control group after anesthesia ( P<0.05), while the level of ROS in HT-22 cells in the isoflurane+ melatonin group was lower than that in the isoflurane group after anesthesia ( P<0.05). (3) There were statistically significant difference in the levels of pDRP1, pAMPK and SIRT1 protein in the hippocampus among the four groups of mice ( F=19.87, 21.20, 25.65, all P<0.05). The levels of pDRP1 and SIRT1 protein in the hippocampus in the isoflurane group were both lower than those in the control group (both P<0.05), while the levels of pDRP1 and SIRT1 protein in the hippocampus in the isoflurane+ melatonin group were both higher than those in the isoflurane group (both P<0.05). In the isoflurane group, the expression of pAMPK protein in the hippocampal region was higher than that in the control group ( P<0.05), while the expression of pAMPK protein in the isoflurane+ melatonin group was lower than that in the isoflurane group ( P<0.05). Conclusion:Melatonin improves long-term isoflurane anesthesia-induced cognitive dysfunction by regulating mitochondrial homeostasis through the AMPK/SIRT1 signaling pathway.

Objective:To explore the medication law of national TCM master Lu Fang in the treatment of primary trigeminal neuralgia (PTN) based on data mining.Methods:With the prescription of the outpatient patients of Harbin Traditional Chinese Medicine Hospital of Professor Lu Fang from September 2014 to September 2022 as the data source, the frequency, property and taste, and meridian tropism of the prescribed drugs were analyzed using Excel 2022 software. R 4.2.1 was used for mining analysis on Chinese materia medica, including correlation, relevance, and clustering,and the medication law in the treatment of PTN was discussed.Results:A total of 300 prescriptions were analyzed, involving 177 kinds of Chinese materia medica, with a frequency of 3 120 times, and 34 kinds of of high-frequency Chinese materia medica. The high frequently Chinese materia medica included Chuanxiong Rhizoma, Angelicae Dahuricae Radix, Puerariae Lobatae Radix, Ligustici Rhizoma et Radix, and Viticis Fructus. The main properties were warm, slightly cold, and neutral, while the main tastes were pungent, bitter, and sweet. The meridian tropism analysis ranked the liver, lung, spleen, and stomach meridians in descending order. Analysis yielded 21 strong association rules, and the association analysis formed a core prescription group based on Chuanxiong Rhizoma, Angelicae Dahuricae Radix, and Ligustici Rhizoma et Radix. The analysis obtained 5 types of clustering combinations.Conclusion:Professor Lu Fang's the medication law to treat primary trigeminal neuralgia is mainly dispelling wind and alleviating pain, which is often combined with the methods, such as searching and dredging collaterals, clearing and dispelling the stagnated heat, calming the liver and subduing yang, soothing the liver and invigorating the spleen.

Objective To investigate shared mechanisms and therapeutic targets between heart failure(HF)and vascular de-mentia(VaD),and identify natural compounds for dual-organ protection.Methods Single-cell data(8 samples)from GEO were processed via Seurat for clustering.Through CellChat,inter-organ communication was constructed,and top 95%ligand-receptor pairs were analyzed by KEGG enrichment.Bulk RNA-seq(70 samples)underwent differential gene(limma)and immune infiltra-tion(CIBERSORT)analyses.31186 compounds from TCMbank werescreened by AutoDock Vina,followed by molecular dynam-ics validation.Results HF fibroblasts(43.75%,7 subclusters)and VaD oligodendrocytes(76.57%,6 subclusters)dominated re-spective tissues.Cross-disease integration revealed HF-driven fibrosis(COLLAGEN)and VaD-associated neuroinflammation(SPP1),converging on PI3K-Akt and ECM pathways.HF-specific markers(TNXB/THBS4/COL1 A2)and VaD signatures(SPP1/PDGFC/TGFA)were identified,with FOXC1 identified as a shared transcriptional regulator.B cell activation character-ized immune dysregulation.Among 8 FOXC1 inhibitors,Qingdainone showed optimal binding[affinity:-9.0 kcal/mol;RMSD:(0.2±0.06)nm].Conclusion This study uncovers fibrosis-neuroinflammation crosstalk in HF-VaD comorbidity and proposes Qingdainone as a FOXC1-targeting therapeutic candidate.

Objective:To investigate the effects and potential mechanisms of melatonin on cognitive dysfunction induced by long-term anesthesia with isoflurane.Methods:Male C57BL/6J mice aged 2 months were divided into control group, isoflurane group, melatonin group, and isoflurane+ melatonin group by random number table, with 6 mice in each group.Three days after anesthesia, cognitive function of mice was assessed by Y-maze and fear conditioning (FC) tests. ATP content in the hippocampus was measured by an ATP assay kit. Western blot was used to detect the expression of DRP1, pDRP1, MFN2, pAMPK and SIRT1 proteins in the hippocampus. Cultured HT-22 cells derived from mouse hippocampal neurons in vitro were divided into control group, isoflurane group, melatonin group, and isoflurane + melatonin group, and the levels of reactive oxygen species (ROS) in each group were detected by flow cytometry after intervention. Statistical analysis was performed by SPSS 22.0 software, and one-way ANOVA was used for comparisons among multiple groups.Results:(1) There was a statistically significant difference in the percentage of freezing behavior in contextual fear memory among the four groups of mice ( F=39.09, P<0.05). The percentage of freezing behavior in the isoflurane group was lower than that in the control group ((44.23±8.88)% vs (75.87±5.90)%, P<0.05), while the percentage of freezing behavior in the isoflurane+ melatonin group((67.45±14.89)%)was higher than that in the isoflurane group ( P<0.05). There was also a statistically significant difference in the percentage of exploration in the novel arm among the four groups of mice ( F=13.87, P<0.05). The percentage of exploration in the novel arm in the isoflurane group was lower than that in the control group((33.64±6.53)% vs (47.13±3.87)%, P<0.05), while the percentage of exploration in the novel arm in the isoflurane+ melatonin group((43.05±1.64)%)was higher than that in the isoflurane group ( P<0.05). (2) Statistically significant difference in the levels of ATP in the hippocampus was found among the four groups of mice ( F=49.22, P<0.05). The level of ATP in the hippocampus in the isoflurane group was lower than that in the control group((2.29±0.15)nmol/mg vs (3.58±0.12)nmol/mg, P<0.05), while the level of ATP in the hippocampus in the isoflurane+ melatonin group ((3.02±0.27)nmol/mg)was higher than that in the isoflurane group ( P<0.05). There was a statistically significant difference in the levels of ROS in HT-22 cells among the four groups ( F=18.36, P<0.05). The level of ROS in HT-22 cells in the isoflurane group was higher than that in the control group after anesthesia ( P<0.05), while the level of ROS in HT-22 cells in the isoflurane+ melatonin group was lower than that in the isoflurane group after anesthesia ( P<0.05). (3) There were statistically significant difference in the levels of pDRP1, pAMPK and SIRT1 protein in the hippocampus among the four groups of mice ( F=19.87, 21.20, 25.65, all P<0.05). The levels of pDRP1 and SIRT1 protein in the hippocampus in the isoflurane group were both lower than those in the control group (both P<0.05), while the levels of pDRP1 and SIRT1 protein in the hippocampus in the isoflurane+ melatonin group were both higher than those in the isoflurane group (both P<0.05). In the isoflurane group, the expression of pAMPK protein in the hippocampal region was higher than that in the control group ( P<0.05), while the expression of pAMPK protein in the isoflurane+ melatonin group was lower than that in the isoflurane group ( P<0.05). Conclusion:Melatonin improves long-term isoflurane anesthesia-induced cognitive dysfunction by regulating mitochondrial homeostasis through the AMPK/SIRT1 signaling pathway.

AIM:To explore the impact of chronic stress on postoperative cognitive dysfunction in rats and to elucidate the mechanistic link to histone deacetylase 2(HDAC2).METHODS:A repeated social defeat stress model and a prolonged isoflurane anesthesia model were established in mice.The rats were randomly assigned to four groups:control(Ctrl)group,isoflurane anesthesia(Iso)group,chronic social defeat stress(RSDS)group,and chronic social de-feat stress combined with isoflurane anesthesia(RSDS+Iso)group.Anxiety-like behaviors were evaluated using the social avoidance test and the novelty-suppressed feeding test.Cognitive function was assessed through the novel object recogni-tion test and the Morris water maze.Plasma corticosterone levels were measured via enzyme-linked immunosorbent assay(ELISA).Primary hippocampal neurons were isolated from fetal mouse hippocampi and classified into four groups:con-trol group,chronic stress combined with prolonged isoflurane anesthesia(Cort+Iso)group,CAY-10683 intervention(CAY),and CAY-10683 treatment(CAY+Cort+Iso)group.Cell viability was determined using CCK-8 assay,and pro-tein expression levels of brain-derived neurotrophic factor(BDNF)and HDAC2 were analyzed by Western blot.RE-SULTS:The RSDS mouse model was successfully established,with ELISA results indicating a significant increase in plasma corticosterone levels in mice subjected to chronic stress combined with prolonged isoflurane anesthesia.Behavioral assessments and Western blot analyses revealed that mice exposed to prolonged isoflurane anesthesia following chronic stress showed marked declines in cognitive function and hippocampal BDNF protein expression levels.Additionally,chronic stress significantly elevated HDAC2 protein expression in the hippocampi of mice undergoing prolonged isoflurane anesthesia.Treatment with an HDAC2 inhibitor reduced HDAC2 protein expression in primary hippocampal neurons sub-jected to chronic stress combined with prolonged isoflurane anesthesia,concurrently increasing BDNF protein expression levels.CONCLUSION:Chronic stress significantly worsens postoperative cognitive dysfunction induced by prolonged isoflurane anesthesia,associated with increased HDAC2 protein expression in the hippocampus.Inhibition of HDAC2 ef-fectively counteracts the reduction in BDNF,a protein crucial for cognitive function,caused by the combination of chronic stress and prolonged isoflurane anesthesia.

Objective To investigate shared mechanisms and therapeutic targets between heart failure(HF)and vascular de-mentia(VaD),and identify natural compounds for dual-organ protection.Methods Single-cell data(8 samples)from GEO were processed via Seurat for clustering.Through CellChat,inter-organ communication was constructed,and top 95%ligand-receptor pairs were analyzed by KEGG enrichment.Bulk RNA-seq(70 samples)underwent differential gene(limma)and immune infiltra-tion(CIBERSORT)analyses.31186 compounds from TCMbank werescreened by AutoDock Vina,followed by molecular dynam-ics validation.Results HF fibroblasts(43.75%,7 subclusters)and VaD oligodendrocytes(76.57%,6 subclusters)dominated re-spective tissues.Cross-disease integration revealed HF-driven fibrosis(COLLAGEN)and VaD-associated neuroinflammation(SPP1),converging on PI3K-Akt and ECM pathways.HF-specific markers(TNXB/THBS4/COL1 A2)and VaD signatures(SPP1/PDGFC/TGFA)were identified,with FOXC1 identified as a shared transcriptional regulator.B cell activation character-ized immune dysregulation.Among 8 FOXC1 inhibitors,Qingdainone showed optimal binding[affinity:-9.0 kcal/mol;RMSD:(0.2±0.06)nm].Conclusion This study uncovers fibrosis-neuroinflammation crosstalk in HF-VaD comorbidity and proposes Qingdainone as a FOXC1-targeting therapeutic candidate.

AIM:To explore the impact of chronic stress on postoperative cognitive dysfunction in rats and to elucidate the mechanistic link to histone deacetylase 2(HDAC2).METHODS:A repeated social defeat stress model and a prolonged isoflurane anesthesia model were established in mice.The rats were randomly assigned to four groups:control(Ctrl)group,isoflurane anesthesia(Iso)group,chronic social defeat stress(RSDS)group,and chronic social de-feat stress combined with isoflurane anesthesia(RSDS+Iso)group.Anxiety-like behaviors were evaluated using the social avoidance test and the novelty-suppressed feeding test.Cognitive function was assessed through the novel object recogni-tion test and the Morris water maze.Plasma corticosterone levels were measured via enzyme-linked immunosorbent assay(ELISA).Primary hippocampal neurons were isolated from fetal mouse hippocampi and classified into four groups:con-trol group,chronic stress combined with prolonged isoflurane anesthesia(Cort+Iso)group,CAY-10683 intervention(CAY),and CAY-10683 treatment(CAY+Cort+Iso)group.Cell viability was determined using CCK-8 assay,and pro-tein expression levels of brain-derived neurotrophic factor(BDNF)and HDAC2 were analyzed by Western blot.RE-SULTS:The RSDS mouse model was successfully established,with ELISA results indicating a significant increase in plasma corticosterone levels in mice subjected to chronic stress combined with prolonged isoflurane anesthesia.Behavioral assessments and Western blot analyses revealed that mice exposed to prolonged isoflurane anesthesia following chronic stress showed marked declines in cognitive function and hippocampal BDNF protein expression levels.Additionally,chronic stress significantly elevated HDAC2 protein expression in the hippocampi of mice undergoing prolonged isoflurane anesthesia.Treatment with an HDAC2 inhibitor reduced HDAC2 protein expression in primary hippocampal neurons sub-jected to chronic stress combined with prolonged isoflurane anesthesia,concurrently increasing BDNF protein expression levels.CONCLUSION:Chronic stress significantly worsens postoperative cognitive dysfunction induced by prolonged isoflurane anesthesia,associated with increased HDAC2 protein expression in the hippocampus.Inhibition of HDAC2 ef-fectively counteracts the reduction in BDNF,a protein crucial for cognitive function,caused by the combination of chronic stress and prolonged isoflurane anesthesia.

Objective:To discuss the improvement effect of uric acid(UA)on the postoperative cognitive dysfunction(POCD)in the mice anesthetized with isoflurane for a long duration,and to clarify its possible mechanism.Methods:Twenty-four healthy male C57BL/6 mice were randomly divided into blank control group,anesthesia group,and UA group,and there were eight mice in each group.The mice in UA group were injected intraperitoneally with 200 μL UA solution daily for 2 d before anesthesia.The mice in blank control group and anesthesia group were given the same volume of saline;the mice in anesthesia group and UA group were used to prepare the models of long-duration isoflurane anesthesia,while the mice in blank control group were untreated.Y-maze tests was used to detect the alternation success rate,movement distances,and movement speeds of the mice in various groups;situational fear experiment was used to detect the percentages of freezing time;Western blotting method was used to detect the expression levels of interleukin(IL)-1β,IL-10,and mature brain-derived neurotrophic factor(mBDNF)proteins in hippocampus tissue of the mice in various groups.Results:The Y-maze test results showed that compared with blank control group,the alternation success rate of the mice in anesthesia group was significantly decreased(P<0.01);compared with anesthesia group,the alternation success rate of the mice in UA group was significantly increased(P<0.01).The situational fear experiment results showed that compared with blank control group,the percentage of freezing time of the mice in anesthesia group was significantly decreased(P<0.01);compared with anesthesia group,the percentage of freezing time of the mice in UA group was significantly increased(P<0.05).The cued memory experiment resutls showed that there were no significant differences of the percentage of freezing time of the mice between various groups(P>0.05).The Western blotting results showed that compared with blank control group,the expression level of IL-1β protein in hippocampus tissue of the mice in anesthesia group was increased(P<0.01),while the expression levels of IL-10 and mBDNF proteins were decreased(P<0.01);compared with anesthesia group,the expression level of IL-1β protein in hippocampus tissue of the mice in UA group was decreased(P<0.05),and the expression levels of IL-10 and mBDNF proteins were increased(P<0.05 or P<0.01).Conclusion:UA can improve the POCD in the mice,and its mechnasim may be related with its anti-inflammatory activity inhibiting the central inflammation and upregulating the mBDNF protein expression.

Alzheimer's disease (AD) is a central neurodegenerative disease with still unclear pathogenesis. Recent studies have shown that axonal transport dysfuction of mitochondria may contribute to AD progression. Normal mitochondrial axonal transport mainly involves microtubules, molecular motors and connexins, while AD early pathological changes can damage mitochondrial axonal transport by interfering with these proteins: accumulated β-amyloid (Aβ) impairs the function of molecular motors; abnormally modified Tau protein reduces microtubule stability; mutant presenilin-1 (PS1) can induce phosphorylation of some related proteins by activating glycogen synthase kinase-3β (GSK-3β); all these processes can damage mitochondrial axonal transport, leading to synaptic dysfunction. This review aims to clarify the possible mechanisms of axonal transport dysfuction of mitochondria in AD and provides new ideas for AD treatment.

Up-frameshift 1 (UPF1), as the most critical factor in nonsense-mediated messenger RNA (mRNA) decay (NMD), regulates tumor-associated molecular pathways in many cancers. However, the role of UPF1 in lung adenocarcinoma (LUAD) amino acid metabolism remains largely unknown. In this study, we found that UPF1 was significantly correlated with a portion of amino acid metabolic pathways in LUAD by integrating bioinformatics and metabolomics. We further confirmed that UPF1 knockdown inhibited activating transcription factor 4 (ATF4) and Ser51 phosphorylation of eukaryotic translation initiation factor 2α (eIF2α), the core proteins in amino acid metabolism reprogramming. In addition, UPF1 promotes cell proliferation by increasing the amino-acid levels of LUAD cells, which depends on the function of ATF4. Clinically, UPF1 mRNA expression is abnormal in LUAD tissues, and higher expression of UPF1 and ATF4 was significantly correlated with poor overall survival (OS) in LUAD patients. Our findings reveal that UPF1 is a potential regulator of tumor-associated amino acid metabolism and may be a therapeutic target for LUAD.
Activating Transcription Factor 4/genetics* ; Adenocarcinoma of Lung ; Amino Acids ; Cell Proliferation ; Eukaryotic Initiation Factor-2 ; Humans ; Lung Neoplasms ; RNA Helicases/metabolism* ; RNA, Messenger/metabolism* ; Trans-Activators/metabolism*