Abstract To understand the relationship between cardiovascular disease (CVD) and adverse childhood experiences (ACEs), the present review aims to describe the burden and influencing factors of CVD, epidemiological characteristics and burden of ACEs, current research on the relationship between ACEs and CVD, and the mechanism of ACEs leading to CVD. It is proposed that further assessment of the relationship is warranted through identifying blood biomarkers, conducting prospective cohort studies and intervention studies. Such efforts would provide valuable scientific insights for primary prevention strategies for cardiovascular disease.

Objective:To investigate the effects of dual vascularized tissue-engineered bone constructed by vascular bundles and endothelial progenitor cells (EPCs) on repair of large bone defects and vascular regeneration.Methods:EPCs were seeded on the demineralized bone matrix (DBM) scaffolds and cultured for 6 days. The attachment and morphology of EPCs on DBM scaffolds were observed by electron microscopy. Next, the radial artery was implanted into a vascular groove opened inside the DBM-EPCs composite scaffolds. Finally, models of a large segmental bone defect were constructed using the radii from 18 New Zealand white rabbits. The rabbits were randomly divided into 4 groups using a simple random sampling method: DBM group, DBM+EPCs group, DBM+vascular bundle group, and DBM+EPCs+vascular bundle group. The DBM group and DBM+EPCs group shared the same rabbits so that transplantations were conducted into the left and right forearms respectively; the DBM+vascular bundle group and DBM+EPCs+vascular bundle group also shared the same rabbits so that transplantations were conducted into the left and right forearms respectively. Consequently, there were 9 experimental sites in each group. X-ray examination and gross morphological observation were performed to evaluate the bone regeneration in the experimental rabbits in each group at 4, 8, and 12 weeks after surgery, and CD31 immunofluorescence staining was used to evaluate the vascular regeneration. Micro-CT was used to analyze bone tissue parameters and reconstruct the three-dimensional structures of the defects site at 12 weeks after surgery.Results:Compared with the DBM, DBM+EPCs and DBM+vascular bundle groups, the DBM+EPCs+vascular bundle group showed new bone tissue crawling on the scaffold surface at 4 weeks after surgery, almost complete healing of the bone defect area at 8 weeks, and forming of a complete and dense bone bridge and appearance of a bone marrow cavity at 12 weeks. Micro-CT data at 12 weeks after surgery showed regular arrangement of the trabeculae, significantly improved mineralization, and increased thickness of the bone cortex in the DBM+EPCs+vascular bundle group. Additionally, in the DBM+EPCs+vascular bundle group, the number of microvessels was significantly higher than that in the other groups at 4, 8, 12 weeks after surgery ( P<0.05), and the angiogenesis and bone tissue regeneration were particularly prominent at 12 weeks after surgery. The number of CD31 cells in the DBM+EPCs+vascular bundle group increased significantly more than that in the DBM, DBM+EPCs and DBM+vascular bundle groups ( P<0.05). Conclusion:As the dual vascularized tissue-engineered bone constructed by vascular bundles and EPCs can significantly promote bone tissue regeneration and angiogenesis, it may be a potential therapeutic strategy for repair of large bone defects.

Objective:To investigate the effects of dual vascularized tissue-engineered bone constructed by vascular bundles and endothelial progenitor cells (EPCs) on repair of large bone defects and vascular regeneration.Methods:EPCs were seeded on the demineralized bone matrix (DBM) scaffolds and cultured for 6 days. The attachment and morphology of EPCs on DBM scaffolds were observed by electron microscopy. Next, the radial artery was implanted into a vascular groove opened inside the DBM-EPCs composite scaffolds. Finally, models of a large segmental bone defect were constructed using the radii from 18 New Zealand white rabbits. The rabbits were randomly divided into 4 groups using a simple random sampling method: DBM group, DBM+EPCs group, DBM+vascular bundle group, and DBM+EPCs+vascular bundle group. The DBM group and DBM+EPCs group shared the same rabbits so that transplantations were conducted into the left and right forearms respectively; the DBM+vascular bundle group and DBM+EPCs+vascular bundle group also shared the same rabbits so that transplantations were conducted into the left and right forearms respectively. Consequently, there were 9 experimental sites in each group. X-ray examination and gross morphological observation were performed to evaluate the bone regeneration in the experimental rabbits in each group at 4, 8, and 12 weeks after surgery, and CD31 immunofluorescence staining was used to evaluate the vascular regeneration. Micro-CT was used to analyze bone tissue parameters and reconstruct the three-dimensional structures of the defects site at 12 weeks after surgery.Results:Compared with the DBM, DBM+EPCs and DBM+vascular bundle groups, the DBM+EPCs+vascular bundle group showed new bone tissue crawling on the scaffold surface at 4 weeks after surgery, almost complete healing of the bone defect area at 8 weeks, and forming of a complete and dense bone bridge and appearance of a bone marrow cavity at 12 weeks. Micro-CT data at 12 weeks after surgery showed regular arrangement of the trabeculae, significantly improved mineralization, and increased thickness of the bone cortex in the DBM+EPCs+vascular bundle group. Additionally, in the DBM+EPCs+vascular bundle group, the number of microvessels was significantly higher than that in the other groups at 4, 8, 12 weeks after surgery ( P<0.05), and the angiogenesis and bone tissue regeneration were particularly prominent at 12 weeks after surgery. The number of CD31 cells in the DBM+EPCs+vascular bundle group increased significantly more than that in the DBM, DBM+EPCs and DBM+vascular bundle groups ( P<0.05). Conclusion:As the dual vascularized tissue-engineered bone constructed by vascular bundles and EPCs can significantly promote bone tissue regeneration and angiogenesis, it may be a potential therapeutic strategy for repair of large bone defects.

Objective To explore the effects of the same dose of fractionated radiation and single radiation on radiation-induced heart damage in mice.Methods Twenty-one wild-type C57BL/6J mice were randomly divided into the normal group,fractionated radiation group and single radiation group.18 Gy X-ray,via fractionated(3 Gy/time,6 times)radiation or single radiation,was used to establish a radiation-induced heart damage model.The concentrations of myocardial enzyme damage markers(creatinekinase(CK),creatinekinase-MB(CK-MB),lactate dehydrogenase(LDH)and LDH1)and peripheral serum ions(K+,Ca2+,Fe2+and Cl-)were detected by an automatic biochemical analyzer at day 7 and 28 after radiation.Ultrasound was used to detect and analyze the cardiac function of mice at day 28 after radiation,including the left ventricular ejection fraction(EF),left ventricular fractional shortening(FS),left ventricular end-diastolic volume(LVEDV),left ventricular mass(LV mass)and left ventricular end-systolic volume(LVESV).The opening of the mitochondrial permeability transition pore(mPTP)and changes of mitochondrial membrane potential of myocardial cells were observed using a laser confocal microscope.The ultrastructure of myocardia was observed under a transmission electron microscope(TEM)and cardiac fibrosis was checked by Masson staining.The atherosclerosis of the aorta was examined by gross oil red staining.Real-time quantitative PCR(RT-qPCR)and Western blotting were performed to detect the expressions ofapoptosis-related genes and proteins,B cell lymphoma-2-associated X protein(BAX)and casepase-3.Results Seven days after 18Gy X-ray irradiation,the expression levels of CK,CK-MB,LDH and LDH1 in the single radiation group increased significantly or trended up,while only CK and LDH1 in the fractionated irradiation group continued to increase.Twenty-eight days after radiation,the expression levels of 4 enzymes in myocardial zymogram were increased by both radiation methods.Seven and twenty-eight days after radiation,the concentrations of serum ions K+,Fe2+,Ca2+and Cl-were significantly decreased by both radiation methods that could lead to the decrease of EF and FS,and the increase of LV mass,LVEDV and LVESV.Single radiation made more difference to EF and FS,and the difference between the two groups was statistically significant.Both methods could decrease the mPTP and mitochondrial membrane potential,especially single radiation,and there was significant difference between the two groups.The results of TEM showed that the mitochondrial cristae of myocardial cells decreased and vacuolated,and the myocardial fiber bundles became thicker after X-ray radiation.Masson staining showed that collagen fibers were deposited in the heart tissue ofmice after X-ray irradiation,particularly in the single radiation group.Gross oil red staining ofthe aorta showed that both methods could damage the aorta of mice,and the area of atherosclerotic plaques in the single radiation group was larger,which was statistically different from that of the fractionated radiation group.The results of RT-qPCR and Western blotting showed that X-ray radiation could increase the expression levels of apoptosis-related BAX and caspase-3 in cardiac tissue,especially in the single radiation group.The mRNAexpressions of BAX and caspase-3 increased more significantly in the single radiation group.Conclusion Both fractionated radiation and single radiation at the same dose can cause heart damage,so they can be used to establish a radiation-induced heart damage model of mice.Single radiation can cause more significant damage to the heart.Different modeling methods can be selected as required.

Objective:To compare the mechanical properties between our self-designed axially controlled compression spinal rod (ACCSR) and conventional spinal rod (CSR) for lumbar spondylolysis (LS).Methods:This study selected 36 ACCSRs (the ACCSR group) and 36 CSRs (the CSR group), both of which were in a diameter of 6.0 mm and manufactured in the same batch. They were subjected respectively to biomechanical tests of spinal rod and pedicle screw-rod internal fixation system. In spinal rod tests: the stiffness and yield load of the spinal rods were calculated using four-point bending tests ( n=7) and comparisons were made between the 2 groups; spinal rod fatigue tests ( n=8) recorded the successful compression loads after 2.5 million cycles of loading and compared them with the maximum force at the isthmus of a normal adult's unilateral lumbar spine (198.72 N). In tests of the pedicle screw-rod internal fixation system, the axial compression tests ( n=7) measured the axial gripping capacity, the axial torsion tests ( n=7) the torsional gripping capacity, and the lateral compression tests ( n=7) the stiffness and yield load of pedicle screws in the 2 groups respectively. Results:The stiffness [(1,543.37±61.41) N/mm] and yield load [1,338.57 (1,282.00, 1,353.80) N] of ACCSR group were significantly smaller than those of CSR group [(3,797.63±156.15) N/mm and 4,059.95 (3,813.80, 4,090.89) N] ( P<0.05). The spinal rod fatigue tests showed that the respective loads of CSR and ACCSR passing the 2.5 million fatigue tests were 640.00 N and 320.00 N, both larger than the maximum force at the unilateral lumbar isthmus of a normal adult (198.72 N). There were no significant differences between the ACCSR group and the CSR group in the axial gripping capacity and torsional gripping capacity, as well as in stiffness and yield load of screws between the 2 groups ( P>0.05). Conclusions:In fixation of LS, although the yield load, stiffness and fatigue resistance of ACCSR are inferior to those of CSR, the biomechanical properties of the two sets of pedicle screw-rod internal fixation system are comparable. The fatigue resistance of ACCSR can meet the stress requirements of the normal human isthmus.

Tissue engineering bone technology, grounded in seed cells, cytokines, and scaffold supports, provides an effective solution for addressing extensive bone defects, demonstrating significant potentials in the field of bone repair. However, this technology still faces numerous challenges. Focusing on vascularization in engineered bones, this article reviews various methods to enhance vascularization within tissue-engineered bones, including multicellular co-culture, application of angiogenic factors, advanced 3D printing, and aid of surgical interventions. This article also analyses the latest research developments and the limitations of the methods, and speculates future research directions for tissue engineered bone.

Objective:To investigate the impact of non-alcoholic fatty liver disease (NAFLD) on the incidence of type 2 diabetes mellitus (T2DM) in an elderly population.Methods:A rural elderly population of Kunshan city, Jiangsu Province were used as the research subject. Prior diabetes mellitus, heavy alcohol consumption and incomplete data were excluded from prospective cohort study analysis. Annual physical examination and follow-up were conducted from 2007 to 2016. T2DM onset, death and loss to follow-up visits were observed as the research subject end points. According to the baseline physical examination results, the study subjects were divided into NAFLD and control groups, and further baseline data of both groups were analyzed whether there were match. The cumulative incidence rate of T2DM were statistically analyzed and compared between the two groups. Simultaneously, the relationship between the two groups of various indexes and the newly developed T2DM were analyzed using Kaplan-Meier. The variables with P < 0.1 were selected and incorporated into the Cox proportional hazard regression model. The impact of NAFLD on the incidence of T2DM was analyzed in an elderly population. Results:At baseline, there were statistically significant differences in the distribution of age, sex, waist circumference, body mass index, systolic blood pressure, diastolic blood pressure, direct bilirubin, blood urea nitrogen and triglycerides between NAFLD and non-NAFLD groups. However, fasting blood glucose, serum creatinine, total bilirubin, total cholesterol, low density lipoprotein cholesterol and high-density lipoprotein cholesterol had no statistically significant differences between the two groups. After nine years of follow-up, 207 newly developed T2DM cases, 52 NAFLD cases and 155 control group were selected as the study subjects. The cumulative incidence rates were 4.25%, 10.34%, and 3.55%, respectively. Kaplan-Meier analysis result showed that there were statistically significant differences in the cumulative incidence rates between the two groups from five-year. NAFLD had increased the T2DM risk in an elderly population by approximately 2.14 times (2.14 CI: 1.132 ~ 4.047) at five-year, and then had increased year by year thereafter. Univariate analysis showed that T2DM risk was 2.76 times higher in NAFLD than non-NAFLD groups (95% CI: 2.015 ~ 3.777). After adjustment for gender, age, body mass index, waist circumference, systolic blood pressure, diastolic blood pressure, serum creatinine, triglyceride, total cholesterol, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol, T2DM risk ratio was 1.68 times higher in NAFLD than control groups (95% CI: 1.163 ~ 2.425). Conclusion:NAFLD is an independent long-term risk factor for the T2DM onset in an elderly population.

A novel paper-based analytical device(PAD)was prepared and applied to determine the xanthine oxi-dase(XOD)inhibitory activity of Salvia miltiorrhiza extracts(SME).First,polycaprolactone was 3D printed on filter paper and heated to form hydrophobic barriers.Then the modified paper was cut according to the specific design.Necessary reagents including XOD for the colorimetric assay were immobilized on two separate pieces of paper.By simply adding phosphate buffer,the reaction was performed on the double-layer PAD.Quantitative results were obtained by analyzing the color intensity with the specialized device system(consisting of a smartphone,a detection box and sandwich plates).The 3D-printed detection box was small,with a size of 9.0 cm x 7.0 cm x 11.5 cm.Color component G performed well in terms of linearity and detection limits and thus was identified as the index.The reaction con-ditions were optimized using a definitive screening design.Moreover,a 10％glycerol solution was found to be a suitable stabilizer.When the stabilizer was added,the activity of XOD could be maintained for at least 15 days under 4℃or-20℃storage conditions.The inhibitory activity of SME was investigated and compared to that of allopurinol.The results obtained with the PAD showed agreement with those ob-tained with the microplate method.In conclusion,the proposed PAD method is simple,accurate and has a potential for point-of-care testing.It also holds promise for use in rapid quality testing of medicinal herbs,intermediate products,and preparations of traditional Chinese medicines.

Objective To understand and analyze the incidence rate,risk factors,independent risk factors and the causes of death in elderly population with non-alcoholic fatty liver disease (NAFLD).Furthermore,analyze the relationship between metabolic syndrome (MS) and mortality rate in patients with NAFLD to provide evidence for the prevention and control of NAFLD in the elderly population.Methods A total of 7 619 elderly people aged over 60 years,and local household registered in Kunshan city,Jiangsu province in 2016 were included as subjects to analyze the incidence rate,influencing factors and causes of death in patients with NAFLD and the relationship between MS and mortality rate in patients with NAFLD.According to different data,using Kruskal-Wallis H test,analysis of variance,t-test,chi-square test or logistic regression analysis were performed.Results The prevalence of NAFLD was14.10％ (1 074/7 619) among the elderly over 60 years in Kunshan city,Jiangsu province in 2016,and the female prevalence rate was significantly higher than that of males (P ＜ 0.05).When the body mass index (BMI) was ＜ 32 (kg/m2),the prevalence of NAFLD increased with the increase of BMI index.When BMI was 18.5-23.9 kg/rn2,24-27.9 kg/m2 and ＞ 32 kg/m2,the prevalence of NAFLD was significantly higher in females than males (P ＜ 0.05).There was no significant difference in prevalence between males and females with BMI ＜18.5 kg/m2 and 28～31.9 kg/m2 (P＞ 0.05).BMI gradually decreased (P ＜ 0.05) with the increase of age,and the incidence of NAFLD showed a downward trend (P ＜ 0.05).An independent risk factors for NAFLD (P ＜ 0.01) were gender (OR =0.616),age (OR =0.970),waist circumference (OR =1.065),triglycerides (OR =1.162),BMI (OR =1.238),and diastolic blood pressure (OR =1.012).The probability of NAFLD combined with three and four kinds of MS was significantly higher than control group (P ＜ 0.05).Subjects' mortality rate during the follow-up period was 1.94％,2.23％ in the NAFLD group,and 1.89％ in the control group.Average life expectancy was 75.58 years in NAFLD group and 78.68years in the control group.All deaths in NAFLD groups were associated with MS,and 70.83％ combined with three or more MS.The primary canse of death in NAFLD group was tumors (37.5％),followed by cardiovascular disease (16.67％) and three cases died directly from MS.Conclusion The prevalence of NAFLD in the elderly population in Kunshan city,Jiangsu province is relatively low,which may be associated with local diet and highintensity workouts.MS disorders represented by obesity and diabetes mellitus are closely associated to the onset of NAFLD.Obesity is the most important risk factor for the incidence of NAFLD in the local elderly population over 60 years old.However,the risk of NAFLD should not be neglected in people with normal BMI,especially in aged women,and the weight control should be the most important means to prevent and control NAFLD.NAFLD may increase mortality rate and reduce life expectancy in the elderly population.The main reason to cause death in the elderly with NAFLD is cardiovascular disease and malignant tumors.Simultaneously,multiple MS development may increase the death rate in elderly with NAFLD.

BACKGROUND:There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned.
OBJECTIVE:To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament
METHODS:We constructed fibroin fiber/smal intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cel s (bone marrow mesenchymal stem cel s) on the scaffold to built cel seeding group. The control group had no treatment. After that, we detected the density of cel adhesion, cel proliferation ability and extracel ular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks.
RESULTS AND CONCLUSION:After 4 hours of incubation, bone marrow seeding group was significantly higher than the cel seeding group in cel adhesion density and proliferation rate (P<0.05). Bone marrow seeding group and cel seeding group showed higher type I, III col agen secretion compared with the control group (P<0.05), but the col agen secretion of bone marrow seeding group and cel seeding group showed no significant difference. Composite cel scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.