1.In Vitro and in Vivo Evaluation of Scutellarin-phospholipid Complex Nanoemulsion and Analysis of Its Activity in Ameliorating LPS-induced Vascular Endothelial Injury
Tian LUO ; Zhiyong HE ; Xiangjun MAO ; Xue LIU ; Jinggang HE ; Yuan ZHI ; Xiangchun SHEN ; Qianli XU ; Ling TAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):159-168
ObjectiveTo evaluate some properties of scutellarin-phospholipid complex nanoemulsion(SCU-PC-NE), such as release, cell uptake and tissue distribution, and to investigate its effect on ameliorating lipopolysaccharide(LPS)-induced vascular endothelial injury. MethodSCU-PC-NE was prepared by weighting SCU-PC, ethyl oleate, Kolliphor HS15, 1,2-propylene glycol(50, 400, 514.3, 85.7 mg), respectively. And the appearance of SCU-PC-NE was observed by transmission electron microscope, the average paticle size and Zeta potential were measured by nanopotential particle size analyzer. The cumulative release of SCU-PC-NE in vitro was measured by dynamic dialysis, thiazolyl blue(MTT) colorimetric assay was used to investigate the effect of SCU-PC-NE on the viability of human umbilical vein endothelial cells(HUVECs), the inverted fluorescence microscope and flow cytometry were used to investigate cell uptake of HUVECs by SCU-PC-NE in vitro using coumarin 6 as a fluorescent probe, the tissue distribution of DiR/SCU-PC-NE labeled by near infrared fluorescent dyes was obeserved by small animal in vivo imaging system. The inflammation injury model was established by co-incubation with LPS(1 mg·L-1) and HUVECs, the effect of SCU-PC-NE on the levels of interleukin(IL)-1β and IL-6 were determined by enzyme-linked immunosorbent assay(ELISA), 18 Kunming male mice were randomly divided into blank group, model group, blank preparation group(equivalent to high dose group), SCU group and SCU-PC-NE low and high dose groups(5, 10 mg·kg-1), 3 mice in each group, and the drug administration groups were administered once in the tail vein at the corresponding dose every 48 h, equal volume of normal saline was given to the blank group and the model group, and the drug was administered for 4 consecutive times. Except for the blank group, the endothelial inflammatory injury was induced by intraperitoneal injection of LPS(10 mg·kg-1) at 12 h before the last administration in each group. Hematoxylin-eosin(HE) staining was used to investigate the effect of SCU-PC-NE on the histopathological changes in the thoracic aorta of mice. ResultThe appearance of SCU-PC-NE displayed pale yellow milky light, mostly spherical with rounded appearance and relatively uniform particle size distribution, with the average particle size of 35.31 nm, Zeta potential of 7.23 mV, and the encapsulation efficiency of 75.24%. The cumulative release in vitro showed that SCU-PC-NE exhibited sustained release properties compared with SCU. The cell viability of SCU-PC-NE was >90% at a concentration range of 1.05-8.4 mg·L-1. The results of cellular uptake experiments showed that the cellular uptake ability of SCU-PC-NE was significantly enhanced when compared with the SCU group(P<0.01). Compared with normal mice, the results of tissue distribution showed that the fluorescence intensity of DiR/SCU-PC-NE was significantly enhanced in the spleen, kidney, brain and thoracic aorta of mice at different time points after intraperitoneal injection of LPS(P<0.05, P<0.01), especially in thoracic aorta. ELISA results showed that the levels of IL-1β and IL-6 in the model group were significantly increased when compared with the blank group(P<0.05, P<0.01), and compare with the model group, all administration groups significantly down-regulated IL-1β level, with the strongest effect in the SCU-PC-NE high-dose group(P<0.01), and all administration groups significantly down-regulated IL-6 level, with the strongest effect in the SCU-PC-NE low-dose group(P<0.05). Compare with the blank group, the results of HE staining showed that the endothelial cells were damaged, the elastic fibers were broken and arranged loosely in the model group, although similar vascular injury could be observed in the blank preparation group, SCU group and SCU-PC-NE low-dose group, the vascular endothelial damage was significantly reduced in the high-dose group of SCU-PC-NE, which had a better effect than that in the SCU group. ConclusionSCU-PC-NE can promote the uptake of drugs by endothelial cells and effectively enriched in the site of vascular endothelial injury caused by LPS, suggesting that it has a protective effect on vascular endothelial injury and is a good carrier of SCU.
2.Establishment of a Multiplex Detection Method for Common Bacteria in Blood Based on Human Mannan-Binding Lectin Protein-Conjugated Magnetic Bead Enrichment Combined with Recombinase-Aided PCR Technology
Jin Zi ZHAO ; Ping Xiao CHEN ; Wei Shao HUA ; Yu Feng LI ; Meng ZHAO ; Hao Chen XING ; Jie WANG ; Yu Feng TIAN ; Qing Rui ZHANG ; Na Xiao LYU ; Qiang Zhi HAN ; Xin Yu WANG ; Yi Hong LI ; Xin Xin SHEN ; Jun Xue MA ; Qing Yan TIE
Biomedical and Environmental Sciences 2024;37(4):387-398
Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannan-binding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP. Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays. Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05). Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.
3.Exploration of the Effect of Guhuaisi Kangfu Pills on Neovascularisation of Steroid-Induced Osteonecrosis of the Femoral Head in Rats Based on Gene Expression of VEGF/PI3K/Akt Pathway
Wen-Xi LI ; Liang-Yu TIAN ; Jin ZHANG ; Cai-Hong SHEN ; Zhi-Min YANG ; Xiao-Yan FENG ; Jia-Qiao GUO ; Yu-Ju CAO
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(8):2127-2135
Objective To observe the therapeutic effect and mechanism of Guhuaisi Kangfu Pills on rats with steroid-induced osteonecrosis of the femoral head(SONFH).Methods Sixty rats were randomly divided into blank group,model group,Xianling Gubao Capsules group and Guhuaisi Kangfu Pills low-,medium-and high-dose groups,10 rats in each group.Except for the blank group,the SONFH model was established by lipopolysaccharide combined with Glucocorticoid induction method in all other groups of rats.At the end of the intervention,for the femoral head,blood vessel radiography was performed to observe the microvascular changes in the bone marrow,and hematoxylin-eosin(HE)staining and calculation of the empty bone trap rate,Micro-CT scanning analysis,and compression experiments were carried out,and the real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the gene expressions of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt)1,vascular endothelial growth factor(VEGF)and platelet endothelial cell adhesion molecule 1(CD31)in whole blood.Results Compared with the blank group,the blood supply in the femoral head medullary cavity of the model group was poor,the empty bone lacuna rate was increased(P<0.05),the bone mineral density and bone volume fraction were significantly decreased(P<0.05),the maximum load and elastic modulus of the femoral head were decreased(P<0.05),and the mRNA expression levels of Akt1,PI3K,VEGF and CD31 in whole blood were decreased(P<0.05).Compared with the model group,the blood supply in the femoral head medullary cavity was relatively good,the empty bone lacuna rate was decreased(P<0.05),the bone mineral density,bone volume fraction,trabecular number and trabecular thickness were significantly increased(P<0.05),the trabecular separation was significantly decreased(P<0.05),the maximum load and elastic modulus of the femoral head were increased(P<0.05),and the mRNA expression levels of Akt1,PI3K,VEGF and CD31 in the whole blood were increased(P<0.05)in the high-dose group of Guhuaisi Kangfu Pills and Xianling Gubao Capsules group.There was no significant difference in the above indexes between the high-dose group of Guhuaisi Kangfu Pills and the Xianling Gubao Capsules group(P>0.05).Conclusion Guhuaisi Kangfu Pills improves SONFH in rats,and its mechanism is related to the promotion of VEGF/PI3K/Akt pathway gene expression,thereby promoting angiogenesis.
4.Study on the Efficacy and Safety of Modified Huanglian Wendan Decoction in Treating Hypertension Complicated with Sleep Disorders of Phlegm-Heat Harassing the Interior Type
Zu-Qiang WU ; Hui-Ping ZHOU ; Jun ZOU ; Tian CAI ; Shun-Zhi YANG ; Shen-Rong HUANG ; Ai-Lan LUO
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(9):2319-2325
Objective To investigate the efficacy and safety of Chinese herbal compound modified Huanglian Wendan Decoction in the treatment of hypertension complicated with sleep disorders of phlegm-heat harassing the interior type.Methods A total of 70 patients with hypertension complicated with sleep disorders of phlegm-heat harassing the interior type were randomly divided into a control group and an observation group,35 cases in each group.The patients in the control group were treated with conventional western medicine,and the patients in the observation group were treated with modified Huanglian Wendan Decoction on the basis of treatment for the control group.The course of treatment covered four weeks.The Clinical Global Impression(CGI)score,traditional Chinese medicine(TCM)syndrome score,Pittsburgh Sleep Quality Index(PSQI)score,blood pressure,and serum levels of homocysteine(Hey),interleukin 6(IL-6),interleukin 10(IL-10)and interleukin 8(IL-8)levels in the two groups were observed before and after treatment.Moreover,the safety of medication was evaluated in the two groups.Results(1)After four weeks of treatment,the CGI score of the observation group was(1.63±0.60)points,which was significantly lower than that of the control group[(2.74±0.82)points],and the difference was statistically significant(P<0.01).(2)After treatment,the TCM syndrome score,systolic blood pressure,diastolic blood pressure and PSQI score of the two groups were significantly decreased compared with those before treatment(P<0.01),and the decrease of TCM syndrome score,systolic blood pressure,diastolic blood pressure and PSQI score in the observation group was significantly superior to that in the control group(P<0.05 or P<0.01).(3)After treatment,the levels of serum Hcy,IL-6 and IL-8 in the two groups were significantly decreased compared with those before treatment(P<0.01),and the serum IL-10 level was significantly increased compared with that before treatment(P<0.01).The decrease of serum Hcy,IL-6 and IL-8 levels and the increase of serum IL-10 level in the observation group were significantly superior to those in the control group(P<0.01).(4)During the treatment,no treatment-related adverse reactions or complications occurred in the two groups.Conclusion Modified Huanglian Wendan Decoction has certain efficacy on patients with hypertension complicated with sleep disorders of phlegm-heat harassing the interior type.The decoction can not only reduce the blood pressure and serum Hcy level of the patients,but also regulate the level of inflammatory factors with high safety.
5.The Identification Idea of Antibodies Against Ku and Other High-Frequency Antigens
Fei-Fei ZHANG ; Jing-Wei LI ; Wei SHEN ; Yi HE ; Hong YUAN ; Li TIAN ; Zhi-Jun YE
Journal of Experimental Hematology 2024;32(3):875-882
Objective:This study was aimed to provide ideas for identifying the antibodies to high-frequency antigens by analyzing a female case of high-frequency antigen antibody(anti-Ku)using serological and sequencing method.Methods:The methods for identification of blood group,erythrocyte antigen,screening and identification of antibody were used to detect the blood type and antibody in the proband.The proband's serum and reagent screening cells treated with Sulfhydryl reagent were applied to judge the type and characteristics of this antibodies when reacted with the regaent screening cells or proband's serum respectively.Gene sequencing was used to determine the genotype of the proband's blood group.Results:The proband's red blood cells were determined as O type RhD positive,whose serum showed strong positive reaction to antibody-screening cells and antibody identification cells with the same intensity in saline and IAT medium,however,the self-cells showed negative effect.The Direct Antihuman Globulin of proband's red blood cells also showed weak positive reaction,and the other blood types were CcEe,Jk(a+b-),P1-,Le(a-b-),Lu(a-b+),K-,k-,Kp(a-b-).Serum of the proband treated with 2-ME still react with three groups of screening cells in IAT medium.The reaction intensity of proband's serum was also unchanged with the cells modified with papain and bromelain,but showed negative effect when the cells were treated with sulfhydryl agents including DTT and 2-ME.Gene sequencing revealed that the KEL genotype of the patient was KEL*02N.24.This patient had a rare K0 phenotype.Conclusion:The rare Kell-null blood group(also known as K0)were identified by serological and molecular tests in the proband who produced both IgG and IgM type of antibody to high-frequency antigen(anti-Ku).These two methods are of great significance in the identification of this rare blood group as well as the antibody to high frequency antigen.
6.Guidelines for management of pediatric acute hyperextension spinal cord injury.
Lian ZENG ; Yu-Long WANG ; Xian-Tao SHEN ; Zhi-Cheng ZHANG ; Gui-Xiong HUANG ; Jamal ALSHORMAN ; Tracy Boakye SEREBOUR ; Charles H TATOR ; Tian-Sheng SUN ; Ying-Ze ZHANG ; Xiao-Dong GUO
Chinese Journal of Traumatology 2023;26(1):2-7
Pediatric acute hyperextension spinal cord injury (SCI) named as PAHSCI by us, is a special type of thoracolumbar SCI without radiographic abnormality and highly related to back-bend in dance training, which has been increasingly reported. At present, it has become the leading cause of SCI in children, and brings a heavy social and economic burden. Both domestic and foreign academic institutions and dance education organizations lack a correct understanding of PAHSCI and relevant standards, specifications or guidelines. In order to provide standardized guidance, the expert team formulated this guideline based on the principles of science and practicability, starting from the diagnosis, differential diagnosis, etiology, admission evaluation, treatment, complications and prevention. This guideline puts forward 23 recommendations for 14 related issues.
Child
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Humans
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Spinal Cord Injuries/complications*
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Spinal Cord
7.Network meta-analysis comparing the clinical outcomes and safety of robotic, laparoscopic, and transanal total rectal mesenteric resection for rectal cancer.
Yuan LIU ; Wei SHEN ; Zhi Qiang TIAN ; Yin Chao ZHANG ; Guo Qing TAO ; Yan Fei ZHU ; Guo Dong SONG ; Jia Cheng CAO ; Yu Kang HUANG ; Chen SONG
Chinese Journal of Gastrointestinal Surgery 2023;26(5):475-484
Objective: To methodically assess the clinical effectiveness and safety of robot-assisted total rectal mesenteric resection (RTME), laparoscopic-assisted total rectal mesenteric resection (laTME), and transanal total rectal mesenteric resection (taTME). Methods: A computer search was conducted on PubMed, Embase, Cochrane Library, and Ovid databases to identify English-language reports published between January 2017 and January 2022 that compared the clinical efficacy of the three surgical procedures of RTME, laTME, and taTME. The quality of the studies was evaluated using the NOS and JADAD scales for retrospective cohort studies and randomized controlled trials, respectively. Direct meta-analysis and reticulated meta-analysis were performed using Review Manager software and R software, respectively. Results: Twenty-nine publications comprising 8,339 patients with rectal cancer were ultimately included. The direct meta-analysis indicated that the length of hospital stay was longer after RTME than after taTME, whereas according to the reticulated meta-analysis the length of hospital stay was shorter after taTME than after laTME (MD=-0.86, 95%CI: -1.70 to -0.096, P=0.036). Moreover, the incidence of anastomotic leak was lower after taTME than after RTME (OR=0.60, 95%CI: 0.39 to 0.91, P=0.018). The incidence of intestinal obstruction was also lower after taTME than after RTME (OR=0.55, 95%CI: 0.31 to 0.94, P=0.037). All of these differences were statistically significant (all P<0.05). There were no statistically significant differences between the three surgical procedures regarding the number of lymph nodes cleared, length of the inferior rectal margin, or rate of positive circumferential margins (all P>0.05). An inconsistency test using nodal analysis revealed no statistically significant differences between the results of direct and indirect comparisons of the six outcome indicators (all P>0.05). Furthermore, we detected no significant overall inconsistency between direct and indirect evidence. Conclusion: taTME has advantages over RTME and laTME, in terms of radical and surgical short-term outcomes in patients with rectal cancer.
Humans
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Robotics
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Robotic Surgical Procedures/adverse effects*
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Network Meta-Analysis
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Retrospective Studies
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Postoperative Complications/etiology*
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Transanal Endoscopic Surgery/methods*
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Rectum/surgery*
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Rectal Neoplasms/pathology*
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Laparoscopy/methods*
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Treatment Outcome
8.Expert consensus on the prevention and treatment of adverse reactions in subcutaneous immunotherapy(2023, Chongqing).
Yu Cheng YANG ; Yang SHEN ; Xiang Dong WANG ; Yan JIANG ; Qian Hui QIU ; Jian LI ; Shao Qing YU ; Xia KE ; Feng LIU ; Yuan Teng XU ; Hong Fei LOU ; Hong Tian WANG ; Guo Dong YU ; Rui XU ; Juan MENG ; Cui Da MENG ; Na SUN ; Jian Jun CHEN ; Ming ZENG ; Zhi Hai XIE ; Yue Qi SUN ; Jun TANG ; Ke Qing ZHAO ; Wei Tian ZHANG ; Zhao Hui SHI ; Cheng Li XU ; Yan Li YANG ; Mei Ping LU ; Hui Ping YE ; Xin WEI ; Bin SUN ; Yun Fang AN ; Ya Nan SUN ; Yu Rong GU ; Tian Hong ZHANG ; Luo BA ; Qin Tai YANG ; Jing YE ; Yu XU ; Hua Bin LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2023;58(7):643-656
9.Analysis of left hepatolithiasis treated with cranial-dorsal approach laparoscopic anatomic left hemihepatectomy
Zhi LIU ; Xianbo SHEN ; Lixue ZHOU ; Jun WANG ; Bingzhang TIAN
Chinese Journal of Hepatobiliary Surgery 2023;29(6):434-437
Objective:To study the safety and feasibility of anatomic left hemihepatectomy via cranial-dorsal approach in the treatment of left hepatolithiasis.Methods:Clinical data of 47 patients with left intrahepatic bile duct stones who underwent cranial-dorsal approach laparoscopic anatomic left hemihepatectomy in Hunan People's Hospital from October 2016 to June 2022 were retrospectively analyzed, including 15 males and 32 females, aged (56.45±1.37) years old. The operative time, intraoperative blood loss, postoperative liver function and complications were analyzed. Patients were followed up by telephone and outpatient review.Results:All 47 patients successfully underwent laparoscopic surgery without conversion. The median operative time was 260 (range, 160-440) min. The median intraoperative blood loss was 100 ml (range, 20-400 ml). The total bilirubin, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were 15.7 (11.7, 21.9) μmol/L, 126.6 (91.7, 168.5) U/L, and 151.1 (98.0, 212.4) U/L on postoperative day (POD) 1, respectively, and decreased to 12.6 (9.6, 16.2) μmol/L, 97.9 (60.7, 156.9) U/L, 54.2 (40.0, 104.1) U/L on POD 3, respectively. The median postoperative hospital stay was 7 (range, 4-24) d. Postoperative abdominal effusion and infection occurred in one patient, and the complication rate was 2.1% (1/47). Postoperative CT review found residual stones in common bile duct in one patient [2.1% (1/47)]. No stone recurrence or death occurred during postoperative follow-up.Conclusion:Anatomic left hemihepatectomy via cranial-dorsal approach is a safe and feasible surgery for the treatment of left hepatolithiasis.
10.Clinical value of arterial stiffness assessment on risk prediction of vascular stiffness in the octogenarian elderly.
Yu Jia WANG ; Zheng Hui GU ; Xue Ping WU ; Zhi Yi FANG ; Tian Hu WANG ; Shan GAO ; Xue YANG ; Xiao Ying SHEN ; Ting Yu ZHOU ; Qi ZHANG ; Jun Xia LI ; Feng CAO
Chinese Journal of Cardiology 2023;51(10):1069-1074
Objective: This study aimed to analyze clinical factors related to arterial stiffening and establish a risk prediction nomogram of arterial stiffening in the octogenarian(≥80 years). Methods: This study was a retrospective cross-sectional study, which enrolled the octogenarian elderly who underwent physical examination and secondary prevention intervention in the outpatient department of Chinese People's Liberation Army General Hospital from April 2022 to August 2022. Clinical data including demographics, biochemical indicators and medical history were collected. Brachial-ankle pulse wave velocity (baPWV) was detected during the clinical visit. Participants were divided into the control group (baPWV≤1 800 cm/s) and vascular sclerosis group (baPWV>1 800 cm/s). The risk factors of arterial stiffness were analyzed by univariate and logistic regression analysis, and the nomogram model was constructed by R programming language. The predictive effect of the nomogram model was evaluated by the receiver operating characteristic curve (ROC). Results: The median age of the 525 participants was 87.0 (82.0, 92.0) years, 504 (96.0%) were male, 82 in the control group, 443 in the vascular sclerosis group. The baPWV, age, systolic blood pressure, mean arterial pressure and diastolic blood pressure were significantly lower in the control group than those in the vascular sclerosis group (all P<0.05). Logistic regression analysis showed that high-density lipoprotein cholesterol, alanine aminotransferase and amylase were protective factors, and alkaline phosphatase and creatinine were risk factors of arterial stiffening (all P<0.05). The combined nomogram model scores including age, mean arterial pressure and the above five laboratory indicators indicated that mean arterial pressure and serum creatinine levels were strongly correlated with vascular sclerosis. The ROC curve suggested that the nomogram model had good prediction ability. Conclusions: Age, mean arterial pressure, high-density lipoprotein cholesterol, alanine aminotransferase, alkaline phosphatase, amylase and creatinine are independently determinants for increased vascular stiffness. The combined prediction model in this study can provide reference for individualized clinical risk prediction of vascular sclerosis in the octogenarian elderly.
Aged, 80 and over
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Humans
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Male
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Aged
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Female
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Ankle Brachial Index
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Vascular Stiffness/physiology*
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Octogenarians
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Retrospective Studies
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Cross-Sectional Studies
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Alanine Transaminase
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Alkaline Phosphatase
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Creatinine
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Sclerosis
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Pulse Wave Analysis
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Risk Factors
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Amylases
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Lipoproteins, HDL
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Cholesterol

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