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 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 evaluate the effect of long-term intake of ω-3 polyunsaturated fatty acids (ω-3 PUFAs) on the activation of hippocampal microglia in a mouse model of postoperative cognitive dysfunction (POCD).Methods:Ninety-six clean-grade healthy male C57BL/6 mice, aged 8 weeks, weighing 18-24 g, were stratified according to body weight and divided into 4 groups ( n=24 each) by a random number table method: control diet group (group C), ω-3 PUFAs group (group ω), control diet plus POCD group (group C+ P) and ω-3 PUFAs plus POCD group (group ω+ P). Mice were fed a special ω-3 PUFAs diet (DHA 0.14 g/100 g, EPA 0.03 g/100 g) for 12 weeks in group ω and group ω+ P, while mice were fed with a control diet for 12 weeks in group C and group C+ P.Tibial fracture procedures were performed under isoflurane anesthesia to develop the POCD model after 12 weeks of feeding.The fear conditioning test and Y maze test were performed on 1st and 3rd days after developing the model.The mice were sacrificed after behavioral tests, and the hippocampal tissues were removed for determination of the contents of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) (by gas chromatography-mass spectroscopy), density of Iba-1 positive microglia (by immunofluorescence staining), and expression of mature brain-derived neurotrophic factor (mBDNF) and precursor brain-derived neurotrophic factor (pro-BDNF) (by Western blot), and contents of interleukin-1beta (IL-1β) and interleukin-6 (IL-6) (by enzyme-linked immunosorbent assay). Results:Compared with group C, the contents of DHA and EPA were significantly increased, the percentage of freezing time in the contextual test was increased, mBDNF/pro-BDNF ratio was increased ( P<0.05), no significant change was found in the rotation accuracy in Y maze test, density of Iba-1 positive microglia and contents of IL-1β and IL-6 in hippocampus ( P>0.05) in group ω ( P<0.05), and no significant change was found in the contents of DHA and EPA ( P>0.05), the percentage of freezing time in the contextual test and accuracy of rotation in Y maze test were decreased on 1st and 3rd days after operation, the density of Iba-1 positive microglia and contents of IL-1β and IL-6 were increased, and mBDNF/pro-BDNF ratio was decreased in group C+ P ( P<0.05). Compared with group C+ P, the contents of DHA and EPA were significantly increased, the percentage of freezing time in the contextual test and accuracy of rotation in Y maze test were increased on 1st and 3rd days after operation, the density of Iba-1 positive microglia and contents of IL-1β and IL-6 were decreased, and mBDNF/pro-BDNF ratio was increased in group ω+ P ( P<0.05). Conclusions:Long-term intake of ω-3 PUFAs can improve cognitive function in a mouse model of POCD, and the mechanism may be related to inhibition of activation of hippocampal microglia, reduction of inflammatory responses, and thus increasing the mBDNF/Pro-BDNF ratio.

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*

The glymphatic system is a fluid dynamics network that is important for maintaining homeostasis of the brain, and it is also a new target for the treatment of various central nervous system diseases. The crucial point regarding research into the glymphatic system is the microhydrodynamics of the cerebrospinal fluid tracer. This review summarizes the emerging technologies, such as magnetic resonance technology, two photon microscopic imaging technology, near infrared fluorescence imaging technology, and transcranial macroscopic imaging, and summarizes its research applications and technical advantages to provide methodological strategies for basic and clinical research on glymphatic system function.

Numerous studies have suggested that liquid-liquid phase separation (LLPS) may be involved in occurrence and progression of neurodegenerative diseases through mediating immune inflammation, transcriptional regulation, protein homeostasis, genomic stability, and oxidative stress, and regulation of LLPS-mediated protein homeostasis has attracted particular attention. Therefore, this paper reviews the research progress of mechanism of protein homeostasis regulation in neurodegenerative diseases in recent years, and discusses the prospect of LLPS related research.

Objective To evaluate the effects of GLYX-13 on cognitive function after long-time isoflurane anesthesia in mice. Methods A total of 192 healthy male C57∕B6J mice, aged 8 weeks, weig-hing 22-25 g, were divided into 4 groups(n=48 each)using a random number table: control group (group C), isoflurane anesthesia group(group I), GLYX-13 group(group G), and isoflurane anesthesia plus GLYX-13 group(group IG). The animals were exposed to 15% isoflurane for 6 h in I and IG groups. GLYX-13 1 mg∕kg was injected via the caudal vein at 2 h before anesthesia in G and IG groups. Novel ob-ject recognition test and contextual fear conditioning test were performed on 1st, 3rd and 7th days after an-esthesia. The expression of 2B subunits-containing NMDA receptor(NR2B)and cyclic adenosine mono-phosphate response element-binding protein(CREB)mRNA in the hippocampus was detected by quantita-tive real-time polymerase chain reaction after the end of behavioral tests on 1st, 3rd and 7th days after anes-thesia. Results Compared with group C, the percentage of time spent in exploring a novel object, dis-crimination index and percentage of freezing time were significantly decreased, and the expression of NR2B and CREB mRNA in the hippocampus was down-regulated in group I(P <005). Compared with group I, the percentage of time spent in exploring a novel object, discrimination index and percentage of freezing time were significantly increased, and the expression of NR2B and CREB mRNA in the hippocampus was up-regulated in group IG(P <005). Conclusion GLYX-13 can significantly improve the cognitive func-tion after long-time isoflurane anesthesia in mice.

Objective To analysis the clinical features of dorsal pancreas agenesis ( DPA) and the associated diabetes, pancreatitis and other congenital organ malformations.Methods Chinese databases of Sinomed, CQVIP and CNKI using the term of short pancreas, pancreas agenesis, bulbar pancreas and dorsal pancreas, and English databases of PubMed using the term of dorsal pancreas agenesis, short pancreas and pancreas hypoplasia were searched.The clinical manifestation, pancreatic head characteristics and associations with diabetes, pancreatitis and other congenital organ malformations were analyzed.Results Six related publications from Chinese databases were searched and 21 patients were included with 2 cases excluded.Sixty-one publications from English database were searched and 71 patients were included.Thus, a total of 91 patients with DPA were analyzed.Abdominal pain was the most common manifestation, which was reported by 61.5% of the patients. 15.3% patients were identified during regular physical examination. Other manifestations including jaundice, fatigue, abdominal discomfort and diabetes were rare.After removing cases with insufficient information, 39 patients (61.9%) carried abnormal pancreatic head.Prevalence of diabetes or impaired glucose tolerance was 56.7% and the percentage of insulin-dependent diabetes in patients with abnormal glycaemia was 47.3%(n=18).20 patients (26.7%) were associated with pancreatitis, including 15 patients (75.0%) with acute pancreatitis, 1 patient (5.0%) with recurrent pancreatitis, and 4 patients (20.0%) with chronic pancreatitis. Thirty-three patients ( 36.2%) suffered other congenital organ malformations, including 21 patients (63.6%) with splenic malformation, 8 patients (24.2%) with heart malformation, and 17 patients (51.5%) with multi-organs malformations like gastrointestinal malformation, azygos vein and inferior cava vena fusion, duodenal and biliary atresia and renal absence.Conclusions The main diagnostic criteria of DPA was the absence of dorsal pancreatic duct.Diabetes was the most common complication followed by pancreatitis.