Objective:To investigate the role of 24-dehydrocholesterol reductase(DHCR24)in doxorubicin-induced senescence-related dysfunction of vascular endothelial cells.Methods:Human umbilical vein endothelial cells(HUVECs)were induced with 0.05 μM doxorubicin for 48 h to establish a stress-triggered premature senescence model.The lentiviral transfection method was employed to achieve DHCR24 overexpression in HUVECs.Cell senescence was evaluated by β-galactosidase staining and Western blot to detect the expression of the senescence-related molecules cyclin-dependent kinase inhibitor 1A(P21)and nicotinamide adenine dinucleotide dependent histone deacetylase 1(SIRT1).Western blot was performed to detect DHCR24 and endothelial nitric oxide synthase(eNOS)expression during endothelial senescence.DAF-FM DA(an NO fluorescent probe)was used to detect intracellular NO production.Results:In the stress-triggered premature senescence model of HUVECs induced by doxorubicin, the expression of the senescence marker P21 was up-regulated( t=19.44, P<0.01), SIRT1 was down-regulated( t=10.10, P<0.01, and the expression of DHCR24 was down-regulated( t=5.946, P<0.01), compared with the control group.Meanwhile, eNOS and NO expression was inhibited( t=11.26, P<0.01; t=10.83, P<0.01).After DHCR24 overexpression, compared with the control stimulation group, the overexpression stimulation group showed that DHCR24( F=72.10, P<0.01)was up-regulated.DHCR24 overexpression alleviated the doxorubicin-induced decrease in eNOS and NO( F=5.797, P<0.05; F=45.12, P<0.01), compared with the control group. Conclusions:DHCR24 may mitigate doxorubicin-induced senescence-related vascular endothelial dysfunction by modulating the eNOS/NO signaling pathway.

Objective:To explore the effects of different concentrations of 2-hydroxybenzylamine(2-HOBA)on atherosclerosis and vascular smooth muscle cell senescence and the underlying mechanisms.Methods:Fourteen apolipoprotein E-deficient(ApoE-/-)mice were used to establish an atherosclerosis model and were divided into two groups(n=7)using the random number method: a high-fat diet(HD)group and a high-fat diet plus 2-HOBA(1 mg/ml)(HD+ HOBA)group.Pulse wave velocity was used to assess vascular stiffness and a treadmill was used to assess exercise endurance.Oil Red O staining was used to detect the size and number of atherosclerotic plaques.Masson staining was used to detect the morphology of collagen fibers and elastic fibers in the plaque, the size of the necrotic core area of the plaque, and the thickness of the fibrous cap.Mouse smooth muscle cells were treated with different concentrations of 2-HOBA(100 μmol/L, 250 μmol/L and 500 μmol/L)to establish an H 2O 2-induced senescence model.Senescence-associated β-galactosidase staining was used to detect cell senescence.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression levels of senescence-related secretory phenotype factors, and Western blot was used to detect the expression levels of senescence-related signaling proteins. Results:Compared with the HD group, the HD+ HOBA group showed that the area and number of aortic atherosclerotic plaques were decreased, and the atherosclerotic plaques were stabilized.In addition, compared with the HD group, vascular stiffness in the HD+ 2-HOBA group decreased by 26%(2.59±0.32 mm/ms vs.3.50±0.28 mm/ms), with a statistically significant difference( P<0.01), and exercise endurance increased by 62%[(143.74±24.25)m vs.(233.50±30.21)m, P<0.01], suggesting that 2-HOBA was able to improve aortic vascular stiffness and exercise endurance in mice.2-HOBA ameliorated H 2O 2-induced vascular smooth muscle cell senescence and decreased the mRNA levels of H 2O 2-induced senescence-associated secretory phenotype factors such as interleukin-1β, interleukin-6, tumor necrosis factor-α and monocyte chemoattractant protein-1.Meanwhile, 2-HOBA also inhibited the expression of p53 and p21, the key signaling factors of senescence. Conclusions:2-HOBA suppresses the development and progression of atherosclerosis through inhibiting oxidative stress-related p53/p21 signaling activation and ameliorating vascular smooth muscle cell senescence and the aging-related inflammatory phenotype.

Objective:To explore alterations of myocardial mechanics in mice models of physiological aging.Methods:Mice aged from 3 weeks to 24 months were chosen(n=50). Conventional echocardiography and layer speckle tracking echocardiography(LS-STE)were applied to evaluate alterations of myocardial mechanics in different ages(each for 10 mise). Pathological myocardial alterations in the aging process were detected using HE and Masson's trichrome staining.Results:Clear age-dependent changes in mice of different ages were observed when assessed using parameters of conventional echocardiography( P<0.05 for all). Values from parameters of LS-STE also showed that longitudinal, circumferential and radial strain in both the endocardium and the epicardium gradually decreased during physiological aging( P<0.05 for all). Compared with the 3-weeks old group, longitudinal strain in the endocardium was significantly reduced in the 14-months old group[(-23.49±2.32) vs.(-17.19±2.28), P<0.05]; Compared with the 3-weeks old group, radial strain, longitudinal strain in the endocardium, longitudinal strain in the epicardium, circumferential strain in the endocardium and circumferential strain in the epicardium were significantly reduced in the 24-months old group[(32.90±5.01) vs.(21.80±5.54), (-23.49±2.32) vs.(-15.44±2.27), (-13.23±3.19) vs.(-7.59±2.21), (-25.93±4.09) vs.(-20.31±4.08), (-9.30±1.92) vs.(-6.01±1.04), P<0.05)]; HE and Masson's trichrome staining of the myocardium indicated increases in the cross-sectional area of cardiomyocytes and fibrosis during the aging process. Conclusions:LS-STE can accurately detect early cardiac dysfunction and pathological myocardial alterations associated with aging in mice and may serve as an effective tool in evaluating anti-myocardial aging.

Objective:To investigate the role and mechanisms of fibulin-1 in senescence-related calcification of rat vascular smooth muscle cells induced by high-concentrationphosphate treatment.Methods:From September 2020 to September 2021, rat primary vascular smooth muscle cells were extracted from the thoracic aorta and abdominal aorta of 10 male SD rats aged 6 to 8 weeks.Phosphate(2.5 mmol/L Pi)was used to stimulate the calcification of vascular smooth muscle cells(VSMCs)in a model of stress-induced senescence-related calcification.Cellular senescence was assessed by SA-β-gal staining.Cellular calcification was determined by alizarin red staining and quantification of calcium deposition.Phenotypic transformation indexes and the expression of fibulin-1 during the process of calcification were detected by Western blot.The expression of fibulin-1 in primary rat vascular smooth muscle cells was knocked down by siRNA, the expression of pSmad3 was detected by immunofluorescence, and the effects of fibulin-1 on phenotypic transformation indexes of smooth muscle cells were detected by Western blot.The cells were cultured with recombinant fibulin-1 while transforming growth factor beta(TGF-β)inhibitor A83-01 and pSmad3 inhibitor SIS3 were also added.The senescence and calcification indexes of smooth muscle cells were detected by Western blot.Results:In the stress-induced aging model with phosphate stimulation of calcification in rat VSMCs, the expression of fibulin-1 was up-regulated( t=11.20, P<0.01), the expressions of MHC and SM22α was down-regulated( t=7.97, P<0.01; t=10.27, P<0.01), and the expression of osteoblastic phenotype markers OPN and Bmp2 and senescence marker P53 was up-regulated( t=4.79, P<0.01; t=9.56, P<0.01; t=14.07, P<0.01). Knockdown of fibulin-1 attenuated the degree of senescence and calcium deposition in VSMCs( t=12.90, P<0.05)and decreased the expression of OPN, Bmp2 and P53( t=5.92, P<0.05; t=10.15, P<0.01; t=8.28, P<0.01), at the same time, and TGF-β and pSmad3 expression was inhibited( t=12.90, P<0.01; t=7.46, P<0.01). After the addition of TGF-β/ smad3 pathway inhibitors, the stimulatory effect of recombinant fibulin-1 on phenotypic transformation and senescence protein expression inVSMCs was significantly reduced( t=4.52, P<0.01; t=9.82, P<0.01; t=3.85, P<0.05). Conclusions:Fibulin-1 can promote aging-related calcification of vascular smooth muscle cells through the TGF-β/smad3 signaling pathway.

Objective:To analyze the composition and functional characteristics of the intestinal microflora in novel coronavirus pneumonia patients with arterial stiffness, in order to provide empirical evidence for rational use of intestinal microecological modulators.Methods:Patients with novel coronavirus pneumonia admitted to our hospital between April 5 and April 19, 2020 were enrolled as research subjects.They were divided into the stiffness group and the control group according to the results of arterial stiffness.Stool samples were collected within 7 days of admission.Intestinal flora DNA was analyzed and entered into a database, shotgun metagenomic sequencing was performed, and bioinformatics analysis was conducted based on sequencing results.Results:A total of 16 patients with novel coronavirus pneumonia were included in this study, including 7 in the stiffness group and 9 in the control group.Brachial-ankle pulse wave velocity and blood pressure were higher in the stiffness group than in the control group( P<0.05). Beta diversity analysis at the phylum level showed that there were significant differences in the composition of the intestinal flora between the two groups( P<0.05). Patients with arterial stiffness had a lower relative abundance than the controls in fecal Holdemanella, Mitsuokella, Deinococcus, Lachnospira, Turicibacter, Butyrivibrio, Sporomusa, and Halanaerobium, species associated with the production of short-chain fatty acids(SCFAs), regulation of energy metabolism, anti-radiation, anti-oxidative stress and anti-inflammatory effects.The Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that the intestinal flora in the control group was mainly involved in the pathways of lipopolysaccharide biosynthesis, metabolism of SCFA and other amino acids, and membrane transport, while the intestinal flora in the stiffness group was mostly concerned with the pathways of amino acid metabolism and DNA damage repair. Conclusions:The use of microbial agents capable of increasing short-chain fatty acids in patients with novel caronavirus infection may contribute to the restoration of intestinal flora homeostasis.

Objective:To evaluate the effect of supervised high-intensity interval training(HIIT)on physical fitness of elderly patients with type 2 diabetes mellitus.Methods:In a prospective randomized controlled study, 47 elderly type 2 diabetes mellitus patients were randomized into either the HIIT group(n=24)or the control group(n=23). All HIIT sessions were conducted under supervision once every other day for 10 weeks.Each session included 40 cycles that consisted of high-intensity training(resting oxygen consumption + 80% oxygen consumption reserve)for 30 seconds and low-intensity training(resting oxygen consumption + 50% oxygen consumption reserve)for 30 seconds.Cardiopulmonary exercise testing, bioelectric impedance analysis and homeostasis model assessment-2(HOMA-2)were used for the measurement of physical fitness, body composition and insulin sensitivity(HOMA-2 IS)before and after intervention. Results:After 10 weeks, peak oxygen uptake(23.6±4.1 ml·kg -1·min -1vs.21.0±4.6 ml·kg -1·min -1, P<0.05), oxygen uptake at the anaerobic threshold(14.1±1.6 ml·kg -1·min -1vs.12.1±2.3 ml·kg -1·min -1, P<0.01), oxygen pulse at the anaerobic threshold(10.7±2.6 ml/min vs.(9.3±1.9)ml/min, P<0.05)and Ln(100·HOMA-2 IS)(4.6±0.4 vs.4.2±0.5, P<0.01)improved in the HIIT group more than in the control group.There were no significant differences in body composition between the two groups( P>0.05). After adjusting for age and body mass index, there was a linear correlation between peak oxygen uptake and Ln(100·HOMA-2 IS)at baseline( r=0.376, P<0.05), but not between changes in peak oxygen uptake and changes in Ln(100·HOMA-2 IS)( r= 0.05, P>0.05). Conclusions:Ten-week HIIT can improve physical fitness of elderly type 2 diabetes mellitus patients.The benefit comes not only from improvement of insulin sensitivity but also from enhancement of heart function.

The population is commonly susceptible to the 2019 novel coronavirus(2019-nCoV), especially the elderly with comorbidities.Elderly patients infected with 2019-nCoV tend to have higher rates of severe illnesses and mortality.Immunoaging is an important cause of severe novel coronavirus pneumonia(NCP)in the elderly.Due to the combination of underlying diseases, elderly patients may exhibit a typical manifestations in clinical symptoms, supplementary examinations and pulmonary imaging, deserving particular attention.The general condition of the elderly should be considered during diagnosis and treatment.In addition to routine care and measures such as oxygen therapy, antiviral therapy and respiratory support, treatment of underlying disease, nutritional support, sputum expectoration, complication prevention and psychological support should also be considered for elderly patients.Based on literature review and expert panel discussion, we drafted the Key Points for the Prevention and Treatment of the Novel Coronavirus Pneumonia in the elderly, aiming to provide help with the prevention and treatment of NCP and the reduction of harm to the elderly population.

Vascular aging refers to degenerative changes in vascular structure and function.Based on speckle-tracking echocardiography, velocity vector imaging can be used to comprehensively evaluate the mechanical characteristics and wall functions in the process of vascular aging and plays an important role in the early detection and treatment of vascular age-related diseases by accurately locating the vessels and three-dimensionally evaluating the velocity, strain and strain rate in different directions.In this article, the application of this technique in the evaluation of vascular aging and related diseases including hypertension, diabetes and atherosclerosis is described.

Objective To investigate the effects and mechanism of advanced glycosylation end products (AGEs)on endothelial cell senescence and endothelial barrier dysfunction.Methods Human umbilical vein endothelial cells (HUVECs)were isolated and cultured.The cells were randomized into three groups:the control group(normal medium),the bovine serum albumin-treated group(BSA control group)and AEGs group(treated with AEGs-BSA).Senescence of HUVECs were detected by senescence-associated β-galactosidase (SA-beta-Gal)staining.The mRNA and protein expressions of senescence-related genes of p53,p21 and p16 in each group were determined by reverse transcription and real-time PCR(RT-qPCR)and Western blot.Reactive oxygen species(ROS)level was determined by dichlorodihdrofluorescence diacetate (DCFH-DA).The transendothelial electric resistance(TER)were measured by endothelial electric resistance meter.The protein levels of myosin light chain kinase (MLCK),phosphorylated myosin light chain (p-MLC),myosin light chain (MLC)were detected by Western blot.Results Compared with the control group and the BSA control group,the AGEs group showed the significantly increased positive rate of senescence-associated SA-beta-Gal staining (67.30 ± 0.75 ％ vs.7.81 ±0.35 ％ and 7.64 ± 0.91％,respectively,P ＜ 0.01)and the expressions of aging-related genes of p53,p21 and p16 were significantly increased (P ＜ 0.05)There was no significant difference in transendothelial electric resistance(TER)between the control group and theBSAgroup(48.0±6.3 Ω· cm2 vs.42.0±7.8 Ω· cm2,P＞0.05),while TER was lower in the AEGs group than in control group and the BSA group[(27.0±4.2)Ω · cm2 vs.(48.0±6.3)Ω · cm2 and (42.0 ± 7.8) Ω · cm2,P ＜0.01].ROS production had no significant difference between the control group and the BSA group[(38.36 ± 8.55) ％ vs.(41.67 ± 6.93) ％,p ＞ 0.05],while was increased in the AEGs group versus control group and the BSA group[(69.31±8.47)％ vs.(38.36±8.55) ％ and (41.67 ± 6.93) ％,P ＜0.05).The protein expression levels of MLK and p-MLC/MLC were higher in the AGEs group than in the control group and the BSA group(P＜0.05).Conclusions AGEs may lead to endothelial cell senescence and endothelial barrier dysfunction by promoting ROS production and oxidative stress,and by regulating MLCK signaling pathway.

Objective To investigate the unusual drug-induced International Normalized Ratio(INR) change in elderly patients with warfarin treatment and its related mechanism. Methods A retrospective analysis was performed in 41 cases of elderly patients with unusual drug-induced INR change from 2011 to 2015 in Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology. Results INR was increased unusually in 37 cases(90.2%) and was decreased unusually in 4 cases(9.8%).Intravenous drugs(80.5%) were prone to cause unusual INR change.Prostaglandin,antifungal drugs,antiarrhythmic drugs and lipid soluble vitamins were the commonly used drugs that induced adverse reaction,accounting for 26.8%,29.3%,29.3% and 9.8%,respectively. Conclusion Unusual drug-induced INR change is not rare in elderly patients with warfarin treatment during hospitalization.When other drugs are prescribed,INR should be measured more frequently and the dose of warfarin should be adjusted promptly.