BACKGROUND: Epoxyeicosatrienoic acids (EETs), which are metabolites of arachidonic acid catalyzed by cytochrome P450 epoxygenase, are degraded into inactive dihydroxyeicosatrienoic acids by soluble epoxide hydrolase (sEH). Many studies have revealed that sEH gene deletion exerts protective effects against diabetes. Vascular calcification is a common complication of diabetes, but the potential effects of sEH on diabetic vascular calcification are still unknown. METHODS: The level of aortic calcification in wild-type and Ephx2-/- C57BL/6 diabetic mice induced with streptozotocin was evaluated by measuring the aortic calcium content through alizarin red staining, immunohistochemistry staining, and immunofluorescence staining. Mouse vascular smooth muscle cell lines (MOVAS cells) treated with β-glycerol phosphate (0.01 mol/L) plus advanced glycation end products (50 mg/L) were used to investigate the effects of sEH inhibitors or sEH knockdown and EETs on the calcification of vascular smooth muscle cells, which was detected by Western blotting, alizarin red staining, and Von Kossa staining. RESULTS: sEH gene deletion significantly inhibited diabetic vascular calcification by increasing levels of EETs in the aortas of mice. EETs (especially 11,12-EET and 14,15-EET) efficiently prevented the osteogenic transdifferentiation of MOVAS cells by decreasing nidogen-2 (NID2) expression. Interestingly, suppressing sEH activity by small interfering ribonucleic acid or specific inhibitors did not block osteogenic transdifferentiation of MOVAS cells induced by β-glycerol phosphate and advanced glycation end products. NID2 overexpression significantly abolished the inhibitory effect of sEH gene deletion on diabetic vascular calcification. Moreover, NID2 overexpression mediated by adeno-associated virus 9 vectors markedly increased insulin-like growth factor 2 (IGF2) and phospho-ERK1/2 expression in MOVAS cells. Overall, sEH gene knockout inhibited diabetic vascular calcification by decreasing aortic NID2 expression and, then, inactivating the downstream IGF2-ERK1/2 signaling pathway. CONCLUSIONS sEH gene deletion markedly inhibited diabetic vascular calcification through repressed osteogenic transdifferentiation of vascular smooth muscle cells mediated by increased aortic EET levels, which was associated with decreased NID2 expression and inactivation of the downstream IGF2-ERK1/2 signaling pathway.
Animals ; Mice ; Vascular Calcification/metabolism* ; Mice, Inbred C57BL ; Epoxide Hydrolases/metabolism* ; Diabetes Mellitus, Experimental/genetics* ; Male ; Gene Deletion ; MAP Kinase Signaling System/genetics* ; Cell Line ; Immunohistochemistry ; Muscle, Smooth, Vascular/metabolism* ; Signal Transduction/genetics* ; Mice, Knockout

Objective To study the effects of Gouteng Jiangya Jieyu Prescription(Uncariae Ramulus cum Uncis,Gastrodiae Rhizoma,Pheretima,Puerariae Lobatae Radix,Salviae Miltiorrhizae Radix et Rhizoma,etc.)on learning and memory ability,hippocampal inflammatory response and autophagy-related protein expression in rats with hypertension complicated with depression(HD).Methods Thirty spontaneously hypertensive rats(SHR)were randomly divided into model group,positive control group(Levamlodipine Besylate 0.45 mg·kg-1+Fluoxetine Hydrochloride 1.80 mg·kg-1)and Gouteng Jiangya Jieyu Prescription high-,medium-and low-dose groups(25.38,12.69,6.34 g·kg-1).Another 6 SD rats were used as blank control group.The SHR rats were intervened by chronic mild unpredictable stress combined with solitary rearing to replicate the HD rat model.At the same time,intragastric administration was given once a day for 6 weeks.The systolic blood pressure and diastolic blood pressure of rat tail artery were measured by non-invasive sphygmomanometer.The learning and memory ability of rats was detected by Morris water maze test.The ultrastructure of hippocampal neurons was observed by transmission electron microscope.The contents of interleukin-1β(IL-1β),IL-18 and IL-10 in hippocampus were detected by ELISA.The expression of autophagy-related proteins Beclin1 and Bcl-2 in hippocampus was detected by immunohistochemistry.The expression of autophagy-related proteins LC3Ⅰ and LC3Ⅱ in hippocampus was detected by Western Blot.Results Compared with the blank control group,the SBP and DBP of the rats in the model group were significantly increased from week 1-6(P＜0.01).The escape latency was significantly prolonged on the third and fourth day(P＜0.01).The first time of crossing the platform was significantly prolonged(P＜0.01),the times of crossing the platform area was significantly reduced(P＜0.05),and the retention time of the platform area was significantly shortened(P＜0.01).The neuronal cell body was obviously swollen,the ridge was destroyed,the nucleus was shrunk,and a large number of autophagosomes appeared;the contents of IL-1β and IL-18 in hippocampus were significantly increased(P＜0.01).The ratio of LC3Ⅱ/LC3Ⅰ protein expression and the expression of Beclin1 protein in hippocampus were significantly up-regulated(P＜0.05,P＜0.01),and the expression of Bcl-2 protein was significantly down-regulated(P＜0.01).Compared with the model group,the SBP of rats in the low-dose group of Gouteng Jiangya Jieyu Prescription was significantly decreased at the weeks 1,3,4,5,6(P＜0.01),and the DBP was significantly decreased at weeks 1,3,4,5(P＜0.05,P＜0.01).The SBP of the rats in the medium-dose group of Gouteng Jiangya Jieyu Prescription was significantly decreased at weeks 1,5,6(P＜0.01),and the DBP was significantly decreased at week 4(P＜0.05).The SBP of rats in the high-dose group of Gouteng Jiangya Jieyu Prescription was significantly decreased in the first week(P＜0.01).The escape latency of rats in the high-and medium-dose groups of Gouteng Jiangya Jieyu Prescription was significantly shortened on the third day(P＜0.05),and the escape latency of rats in the high-and low-dose groups of Gouteng Jiangya Jieyu Prescription was significantly shortened on the fourth day(P＜0.05).The first crossing platform time of rats in the high-,medium-and low-dose groups of Gouteng Jiangya Jieyu Prescription was significantly shortened(P＜0.01).The times of rats crossing the platform area in the medium-and low-dose groups of Gouteng Jiangya Jieyu Prescription were significantly increased(P＜0.05),and the retention time in the platform area was significantly prolonged(P＜0.05).In the administration group,the degree of hippocampal neuron damage was reduced,the nuclear shrinkage was significantly improved,and the autophagosomes were reduced.The contents of pro-inflammatory factors IL-1β and IL-18 in the hippocampus of rats in the high-and medium-dose groups of Gouteng Jiangya Jieyu Prescription were significantly decreased(P＜0.05,P＜0.01).The content of anti-inflammatory factor IL-10 in the hippocampus of rats in the high-dose group of Gouteng Jiangya Jieyu Prescription was significantly increased(P＜0.01).The protein expression ratio of LC3Ⅱ/LC3Ⅰ in hippocampus of high-,medium-and low-dose groups of Gouteng Jiangya Jieyu Prescription was significantly down-regulated(P＜0.01),and the expression of Bcl-2 protein was significantly up-regulated(P＜0.01).The expression of Beclin1 protein in the hippocampus of the high-and medium-dose groups of Gouteng Jiangya Jieyu Prescription was significantly down-regulated(P＜0.05,P＜0.01).Conclusion Gouteng Jiangya Jieyu Prescription can reduce the tail arterial pressure of HD rats,improve their learning and memory ability,and alleviate hippocampal neuronal damage.The mechanism may be related to reducing the release of inflammatory factors,increasing the level of anti-inflammatory factors,and regulating the expression of hippocampal autophagy-related proteins LC3Ⅱ/LC3Ⅰ,Beclin1 and Bcl-2.

Objective To observe the value of spectral CT for predicting regional lymph node metastasis of colorectal cancer(CRC).Methods Totally 73 patients with CRC confirmed by postoperative pathology were retrospectively enrolled and divided into positive group(n=29)and negative group(n=44)based on the presence or absence of regional lymph node metastasis.The lymph node short diameter,CT value of 120 kV conventional image(CT value120 kV),CT values of 40 keV and 70 keV virtual single energy images(CT value40 keV and CT value70keV),iodine density(ID),effective atomic number(Zeff)and spectral curve slope(λHU).The above parameters and clinical indicators were compared between groups.After excluding those with variance inflation factor greater than 10,model 1 was constructed based on carbohydrate antigen 19-9(CA19-9)and carcinoembryonic antigen(CEA),model 2 based on conventional CT parameters(lymph node short diameter,CT value120 kV),model 3 based on spectral CT parameters(CT value70 keV,ID,Zeff,λHU),and model 4 based on conventional CT parameters and spectral CT parameters.The efficacy of 4 models for predicting regional lymph node metastasis of CRC were analyzed.Results Significant differences in patients'gender,carbohydrate antigen 19-9(CA1 9-9),carcinoembryonic antigen(CEA),lymph node short diameter,CT value120 kV,CT value40 keV,CT value70 keV,ID,Zeff and λHU were found between 2 groups(all P＜0.05).The area under the curve(AUC)of model 1-4 for predicting regional lymph node metastasis of CRC was 0.734,0.752,0.996 and 0.995,respectively,and significant differences were found between model 1 and 3,model 1 and 4,model 2 and 3,as well as model 2 and 4(all P＜0.001).Conclusion Enhanced venous phase spectral CT could effectively predict regional lymph node metastasis of CRC.

With the rapid development of social economy， the number of patients with nervous system diseases has increased， and the incidence of the population has a trend of younger， which has a serious impact on life health and social economy. Artemisinin is an active antimalarial component extracted and isolated from Artemisia annua， a Chinese medicinal material. Artemisinin and its derivatives， in addition to the antimalarial effect， also have anti-parasitic， anti-fungal， anti-viral， hypoglycemic， hypolipidemic， anti-tumor， and anti-inflammatory effects， showing a wide range of pharmacological activities. In the past five years， research on the new pharmacological effects of artemisinin and its derivatives has been deepening， and the efficacy of artemisinin and its derivatives in nervous system diseases has attracted much attention， including anti-neuroinflammation， anti-oxidative stress， maintaining the stability of the blood-brain barrier， regulating the release of neurotransmitters， repairing neuronal damage， and promoting neuronal regeneration. These pharmacological effects indicate that artemisinin and its derivatives are potentially capable of neuroprotection. By sorting out literature on the pharmacological activity of artemisinin and its derivatives in nervous system during 2019－2024， this paper systematically summarized the protective effects of artemisinin and its derivatives against nervous system diseases such as stroke， neurodegenerative diseases， neuroimmunological diseases， neuralgia， and nervous system tumors. This review is expected to provide clues and evidence for new indication expansion of artemisinin drugs， innovative drug development， and clinical treatment of nervous system diseases.

The impact of air pollution on human health has always been a research hotspot in the global health field. Outdoor air pollutants composed of multiple components can enter the human body through various pathways. Cardiovascular diseases are a group of diseases caused by outdoor air pollutants. Studies have shown that the incidence of cardiovascular diseases， including hypertension， arrhythmia， and heart failure， is significantly increased among people exposed to air pollution environments. Air pollutants such as fine particulate matter， nitrogen dioxide， ozone， carbon monoxide， and sulfur dioxide are closely related to the occurrence of cardiovascular diseases， and short-term and long-term exposure causes different cardiovascular risks. By reviewing the relevant research reports from 2019 to 2024， this article summarizes the epidemiological evidence of cardiovascular diseases caused by different air pollutants. It generalizes the pathways through which air pollutants accelerate the progression of cardiovascular diseases. These pathways include oxidative stress， inflammatory response， thrombosis， extracellular vesicle release， endoplasmic reticulum stress， apoptosis， endothelial dysfunction， autonomic nervous system imbalance， and their interactions. Based on the different mechanisms of air pollution on cardiovascular diseases， the article analyzes the main progress in drug intervention and summarizes the roles of various active ingredients and compound prescriptions of traditional Chinese medicine in treating air pollution-related cardiovascular diseases， providing reference for the research on the mechanisms and drug interventions of air pollution-related cardiovascular diseases.

Clustered regularly interspaced short palindromic repeats（CRISPR）/CRISPR associated nuclease 9 （CRISPR/Cas9） is a self-defense system found in bacteria and archaea that enables targeted gene editing based on the principle. Due to its universality， efficiency， and simplicity， CRISPR/Cas9 has been applied in the pathological mechanism and prevention and treatment of diseases in many fields. Cerebrovascular diseases and central nervous system diseases seriously endanger human health. Stroke is related to genetics， unhealthy living habits， chronic diseases， and other factors. The brain tissue structure is complex and the cell types are diverse. It is difficult for a universal gene editing platform to study target genes safely， specifically， and efficiently. Scholars have continuously improved and optimized gene editing technology， explored the potential and research methods of gene editing technology， and promoted the research process of brain science. After a brief introduction to the mechanism of CRISPR/Cas9， this paper mainly summarized the optimization of the system in the fields of cerebral science including delivery methods， adeno-associated virus assembly， and new nanomaterials. Its application in cerebrovascular research including vascular homeostasis， microglial homeostasis， angiogenesis， blood-brain barrier， and drug screening was also summarized. Finally， this paper prospected the development of CRISPR/Cas9 in traditional Chinese medicine， hoping to provide references for related research design.

Objective:To explore the chain mediating effect of future orientation and academic engagement between perceived teacher support and academic procrastination in high school students.Methods:From December 2021 to February 2022, a survey was conducted on 550 high school students by the perceived teacher support questionnaire, the adolescent future orientation scale, the academic engagement scale, and the general procrastination scale-for student populations (GPS). Data were entered by EpiData 3.1 software, and SPSS 26.0 software was used to process and analyze the data by one-way ANOVA, Pearson correlation analysis and Bootstrap method test.Results:The scores of perceived teacher support, future orientation, academic engagement and academic procrastination were (3.77±0.80), (3.22±0.48), (68.11±18.08) and (52.32±11.78) respectively.The results of correlation analysis showed that academic procrastination was negatively correlated with perceived teacher support, future orientation and academic engagement ( r=-0.32, -0.38, -0.49, all P<0.01), while perceived teacher support was positively correlated with future orientation and academic engagement ( r=0.40, 0.43, both P<0.01). Future orientation was positively correlated with academic engagement ( r=0.56, P<0.01). The mediating effect analysis showed that perceived teacher support had a significant direct effect on academic procrastination (effect value: -0.10, 95% CI =-0.19--0.02), accounting for 32.26% of the total effect.The mediating effect between perceived teacher support and academic procrastination was found between future orientation and academic engagement (effect value: -0.05, 95% CI =-0.09- -0.02; effect values: -0.09, 95% CI=-0.15--0.05), accounting for 16.13% and 29.03% of the total effect respectively.Future orientation and academic engagement had a chain mediating effect between perceived teacher support and academic procrastination (effect value: -0.07, 95% CI=-0.10--0.04), accounting for 22.58% of the total effect. Conclusion:Perceived teacher support can influence academic procrastination, not only through the direct path, but also through the indirect path of future orientation and academic engagement, as well as chain mediating path between these two variables.

Objective To investigate the effect of verbascoside(VB)on endothelial dysfunction in atherosclerotic(AS)rats by regulating high-mobility group protein B1(HMGB1)/receptor for advanced glycation endproducts(RAGE)/nuclear factor κB(NF-κB)signal pathway.Methods The rat model of AS was established by high fat feeding combined with vitamin D3 solution intraperitoneal injection.Rats were divided into the control group(n=10),the model group(n=12),the low(VB-L),medium(VB-M)and high dose(VB-H)VB groups(2,5 and 10 mg/kg,n=10),and the positive control group(simvastatin,5 mg/kg,n=10).The serum level of blood lipids was detected by automatic biochemical analyzer.Pathological changes of aorta were observed by HE staining.Serum levels of inflammatory factors and vascular endothelial cytokines were detected by enzyme-linked immunosorbent assay(ELISA).The level of oxidative stress in rats was detected by micro-method kit.The expression of HMGB1/RAGE signal pathway protein in aorta was detected by Western blot assay.Results Compared with the control group,the intima of aorta in the model group was thickened,plaque appeared in blood vessels,accompanied by lipid deposition and inflammatory cell infiltration.Serum levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL-C),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),C-reactive protein(CRP),matrix metalloproteinase-9(MMP-9),endothelin-1(ET-1),visfatin,intercellular adhesion molecule-1(ICAM-1)and malondialdehyde(MDA),and HMGB1,RAGE and phosphorylation levels of NF-κB in aorta were obviously increased.Serum levels of high-density lipoprotein cholesterol(HDL-C),nitric oxide(NO),superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were obviously decreased(P＜0.05).Compared with the model group,pathological changes of rats were obviously improved in the VB-L,VB-M and VB-H groups and the simvastatin group.Serum levels of TC,TG,LDL-C,TNF-α,IL-1β,CRP,MMP-9,ET-1,visfatin,ICAM-1,MDA,and HMGB1,RAGE,phosphorylation levels of NF-κB in aorta were obviously decreased,and serum levels of HDL-C,NO,SOD and GSH-Px were obviously increased(P＜0.05).Conclusion VB can down-regulate the expression of HMGB1/RAGE/NF-κB signal pathway protein,inhibit inflammation and oxidative stress in AS rats,and improve lipid metabolism and vascular endothelial function.

Objective:To explore the value of MR elastography (MRE) and shear wave elastography (SWE) for staging liver fibrosis in a rabbit model.Methods:From March to November 2020, 200 healthy New Zealand white rabbits were randomly divided into control group ( n=40) and liver fibrosis group ( n=160) by random number table method. The volume ratio of CCl 4 and olive oil was 1∶1 to prepare 50% CCl 4 oil solution, and the experimental rabbits in the liver fibrosis group were subcutaneously injected with 50% CCl 4 olive oil solution. It was injected once a week at the dose of 0.1 ml/kg in the first to third weeks, once a week at the dose of 0.2 ml/kg in the 4th to 6th weeks. The dose of 0.1 ml/kg was injected twice a week from week 7 to 16. The control group were subcutaneously injected with an equal dose of normal saline. At the end of the 4th, 8th, 12th, and 16th week, 40 and 10 animals in the liver fibrosis group and the control group were randomly selected by random number table method for MRE and SWE, respectively, to obtain the liver elastic stiffness (LS), which were recorded as LS MRE and LS SWE. After the examination, the experimental rabbits were sacrificed and liver tissue of rabbits were taken for histopathological Scheuer staging, and they were divided into F0-F4 groups. One-way ANOVA was used to evaluate the differences of LS MRE and LS SWE in different stages of liver fibrosis. Spearman correlation was used to analyze the correlation between LS and pathological stages. The receiver operating characteristic curve was used to evaluate the efficacy of LS MRE and LS SWE in diagnosing liver fibrosis staging, and the area under the curve (AUC) was compared using the Z test. Results:Totally 162 rabbits were included, which covered F0 ( n=38), F1 ( n=33), F2 ( n=35), F3 ( n=31) and F4 ( n=25). Significant differences of LS MRE and LS SWE values were found among different stages of liver fibrosis ( F=295.29, 102.40, both P<0.001). LS MRE, LS SWE were both positively correlated with liver fibrosis stage ( r=0.93, 0.81, both P<0.001). The AUC of LS MRE for diagnosing liver fibrosis stages ≥F1, ≥F2, ≥F3, and ≥F4 were 0.955, 0.967, 0.996, and 0.980, respectively; the AUC of LS SWE were 0.856, 0.880, 0.974, and 0.953, respectively. The AUC of liver fibrosis stage ≥ F1, ≥ F2 for LS MRE value were greater than LS SWE value ( Z=2.93, 3.29, P=0.003, 0.001), and the AUC of ≥F3, ≥F4 had no significant differences ( Z=1.58, 1.68, P=0.115, 0.093). Conclusion:Both MRE and SWE can accurately predict the stage of liver fibrosis in experimental rabbits, and MRE is better than SWE in diagnosing early liver fibrosis.