二维斑点追踪显像技术评估窒息新生儿左心室纵向收缩功能与心肌损害的相关性

【摘要】目的 探讨二维斑点追踪显像技术(2D-STI)评估新生儿窒息合并心肌损害后左心室整体及局部心肌的纵向收缩功能在早期诊断窒息新生儿心肌损害中的临床价值。方法 选择2019年07月至2020年12月期间在右江民族医学院附属医院新生儿科住院的足月窒息新生儿61例，经临床确诊合并心肌损害，根据Apgar评分分为轻度组31例和重度组30例，选择同期住院出生的正常足月新生儿30例作为对照组。检测受检者的血清肌酸激酶同工酶（CK-MB）、肌钙蛋白（cTnT）、左室舒张期前后径（LVDId）、左室射血分数（LVEF）、左室短轴缩短率（LVFS）、辛普森法左室射血分数（Simpson EF）、左室三腔心整体应变（GLS-LAX）、左室四腔心整体应变（GLS-A4C）、左室两腔心整体应变（GLS-A2C）、左室整体应变（GLS-AVG）,分析GLS-AVG和CK-MB、cTnT三者的相关性。结果 三组间CK-MB和cTnT比较差异有统计学意义（P<0.05）。三组间性别、体重、胎龄均无统计学差异（P>0.05）。三组间LVDId、LVEF、LVFS、Simpson EF比较差异无统计学意义（P>0.05）。GLS-AVG与CK-MB呈负性相关（r=-0.515，P=0.000），GLS-AVG与cTnT呈负性相关（r=-0.912，P=0.000）。结论 GLS-AVG与CK-MB、cTnT具有相关性，GLS-AVG可作为窒息新生儿心肌损害早期诊断指标。     

阿克替苷通过激活血红素加氧酶-1抑制H_(2)O_(2)诱导的视网膜神经节细胞氧化损伤北大核心

目的研究阿克替苷对视网膜神经节细胞(RGC-5)的保护作用及其机制。方法将RGC-5细胞分为对照组、H_(2)O_(2)组、阿克替苷+H_(2)O_(2)组(1μmol·L^(-1)、10μmol·L^(-1)、30μmol·L^(-1)阿克替苷分别预处理细胞2 h),MTT法检测各组细胞活力;后续实验阿克替苷+H_(2)O_(2)组选择阿克替苷30μmol·L^(-1)作为治疗浓度,荧光探针H2DCF-DA检测活性氧(ROS)产生和罗丹明123检测线粒体膜电位,Western blot检测各组细胞中Bcl-2、Bcl-2/Bax、半胱氨酸蛋白酶-3(Caspase-3)蛋白表达。机制研究中首先用不同浓度阿克替苷单独处理RGC-5细胞24 h,Western blot检测血红素加氧酶-1(HO-1)蛋白表达水平,进一步锌原卟啉(ZnPP)预处理RGC-5细胞1 h,而后30μmol·L^(-1)阿克替苷单独预处理后与200μmol·L^(-1)H_(2)O_(2)共孵育24 h,通过MTT法测定细胞活力。结果与对照组相比,H_(2)O_(2)组中细胞活力明显降低(P<0.01)。与H_(2)O_(2)组相比,阿克替苷+H_(2)O_(2)组中10μmol·L^(-1)、30μmol·L^(-1)阿克替苷预处理可显著提高细胞活力(P<0.05、P<0.01)。与对照组相比,H_(2)O_(2)组RGC-5细胞ROS生成明显增加,线料体膜电位显著降低,同时Bcl-2/Bax比值降低和Caspase-3蛋白相对表达量增加(均为P<0.05)。与H_(2)O_(2)组相比,阿克替苷+H_(2)O_(2)组RGC-5细胞ROS生成减少,线粒体膜电位增加,Bcl-2/Bax比值提高,Caspase-3蛋白相对表达量减少(均为P<0.05)。与阿克替苷未处理组相比,阿克替苷剂量依赖性诱导HO-1蛋白的表达,与阿克替苷+H_(2)O_(2)组相比,加入ZnPP后细胞活力降低(P<0.01),说明HO-1抑制剂ZnPP降低了阿克替苷对H_(2)O_(2)损伤的抑制作用。结论阿克替苷能够通过上调HO-1表达抑制H_(2)O_(2)诱导的RGC-5氧化应激损伤。     

Moderately increased alcohol consumption is associated with higher pressure wave reflections and blood pressure in men

Background and aimsIncreased alcohol consumption has been associated with CVD risk. Subclinical arterial damage (SAD) precedes the onset of cardiovascular disease (CVD), and allows early identification and study of the pathophysiology of CVD. Reliable, noninvasive vascular biomarkers are available for the early detection of SAD and reclassification of CVD risk. To investigate the association of alcohol consumption with multiple SAD biomarkers and central hemodynamics in a large sample of Greek adults with CVD risk factors.Methods and resultsThis cross-sectional study was conducted with 938 participants (43.5% men) and collected data on SAD biomarkers, central hemodynamics, and dietary intake. Multiple linear regression analysis was performed according to sex after adjusting for several confounders. In men, alcohol consumption of 20–30 g/d was positively associated with mean, diastolic, and peripheral systolic blood pressure (BP). The consumption of >30 g/d was positively associated with the augmentation index. In women, no statistically significant associations were found between alcohol consumption and BP or SAD indices. No statistically significant associations were found between alcohol consumption and arterial compliance or distensibility in both sexes.ConclusionIn men even a small deviation from the current recommendation for alcohol consumption is associated with both higher BP indices and pressure wave reflections. The absence of association in women might be due to very low alcohol intake, even in the high consumption group. More studies are needed to verify our findings and establish the above associations in each sex.     

Antitumor effects of dioscin in A431 cells via adjusting ATM/p53-mediated cell apoptosis,DNA damage and migration

Skin cancer is the deadliest type of malignant disease and causes primary mortality worldwide. Dioscin, which exists in medicinal plants, has potent anticancer effects. However, its effects on skin cancer remain unknown. In the present study, the activity and mechanism of dioscin on the human skin cancer A431 cell line were investigated, MTT, colony formation, Transwell, wound-healing, TUNEL, Comet, immunofluorescence and western blot assays were used to assess the effects of dioscin on A431 cells. The results of MTT, colony formation, Transwell and wound-healing assays revealed that dioscin suppressed proliferation, colony formation and invasion of the cancer cells. TUNEL and comet assays demonstrated that dioscin exhibited significant effects on cell apoptosis and DNA damage. Investigations into the mechanism revealed that the expression levels of phosphorylated Ataxia telangiectasia-mutated (ATM) were considerably activated by dioscin, which significantly upregulated the expression levels of p53 to activate mitochondrial apoptosis signaling. Furthermore, the expression levels of BAX, cleaved caspase-3/9 and cleaved poly (ADP-ribose) polymerase were upregulated, and the expression levels of BCL-2 were downregulated by dioscin. Additionally, dioscin markedly downregulated the expression levels of matrix metalloproteinase 2 (MMP2), MMP9, RHO and cdc42, which are all associated with tumor invasion. In addition, p53-small interfering RNA transfection experiments indicated that dioscin exhibited excellent activity against skin cancer in vitro by decreasing p53 expression. Overall, the present results suggested that dioscin inhibited skin cancer cell proliferation via adjusting ATM/p53-mediated cell apoptosis, migration and DNA damage, which should be considered as a potential option for future treatments of skin cancer.     

Therapeutic effectiveness of a single exercise session combined with Walk Aide functional electrical stimulation in post-stroke patients:a crossover design study

A growing body of evidence has suggested that the imbalance of epigenetic markers and oxidative stress appears to be involved in the pathophysiology and progression of stroke. Thus, strategies that modulate these biomarkers might be considered targets for neuroprotection and novel therapeutic opportunities for these patients. Physical exercise has been reported to induce changes in these epigenetic markers and improve clinical outcomes in different populations. However, little is reported on this in post-stroke patients. The purpose of this study was to investigate the effect of a single exercise session with Walk Aide functional electrical stimulation(FES) on cognitive performance, clinical functional parameters, oxidative stress and epigenetic modulation in post-stroke individuals. In this crossover design study, 12 post-stroke individuals aged 54–72 years of either sexes were included and subjected to a single session of exercise(45 minutes) without Walk Aide functional electrical stimulation(EXE alone group), followed by another single session of exercise(45 minutes) with Walk Aide functional electrical stimulation(EXE + FES group). The clinical functional outcome measures, cognitive performance and blood collections for biomarker measurements were assessed pre-and post-intervention. After intervention, higher Berg Balance Scale scores were obtained in the EXE + FES group than in the EXE alone group. There was no significant difference in the Timed Up and Go test results post-intervention between EXE alone and EXE + FES groups. After intervention, a better cognitive performance was found in both groups compared with before the intervention. After intervention, the Timed Up and Go test scores were higher in the EXE + FES group than in the EXE alone group. In addition, the intervention induced lower levels of lipid peroxidation. After intervention, carbonyl level was lower, superoxide dismutase activity and superoxide dismutase/catalase activity ratio were higher in the EXE + FES group, compared with the EXE group alone. In each group, both histone deacetylase(HDAC2) and histone acetyltransferase activities were increased after intervention compared with before the intervention. These findings suggest that a single exercise session with Walk Aide FES is more effective on balance ability and cognitive performance compared with conventional exercise alone in post-stroke patients. This is likely to be related to the regulation of oxidative stress markers. The present study was approved by the Research Ethics Committee of the Methodist University Center-IPA(approval No. 2.423.376) on December 7, 2017 and registered in the Brazilian Registry of Clinical Trials—Re BEC(RBR-9 phj2 q) on February 11, 2019.     

大黄素治疗代谢相关脂肪性肝病的作用机制研究进展

代谢相关脂肪性肝病既往称为非酒精性脂肪肝，与肥胖、糖尿病、高脂血症等关系密切，还明显增加心血管死亡的风险。大黄素是大黄、何首乌中主要的活性成分，具有多种生物学功能。大黄素可通过降低肝细胞脂质沉积、抑制肝脏炎症反应、抗胰岛素抵抗、抑制肝纤维化、抗氧化应激反应等多种途径对代谢相关脂肪性肝病发挥治疗作用。总结了大黄素治疗代谢相关脂肪性肝病的作用机制，为代谢相关脂肪性肝病的治疗提供参考。     

人脐带间充质干细胞对大鼠视网膜光损伤的保护作用

目的　观察体外人脐带间充质干细胞（human umbilical cord mesenchymal stem cells，hUCMSCs）能否由视网膜匀浆上清液诱导分化为神经样细胞及移植到视网膜光损伤大鼠玻璃体内后存活、迁移、整合及分化的情况。方法　无菌条件下采集健康足月剖宫产胎儿的正常脐带组织，经2.5 g·L^-1胰蛋白酶和1 g·L^-1胶原酶消化、贴壁培养获得hUCMSCs。将CM-Dil标记的hUCMSCs注入光损伤大鼠玻璃体内，观察眼内整合分化情况，对正常对照组、光损伤组、PBS治疗组和hUCMSCs治疗组大鼠视网膜外核层层数和厚度进行对比分析。结果　hUCMSCs培养48 h后贴壁生长，呈长梭形，14 d后可见细胞融合成片。光损伤后大鼠视网膜外核层结构紊乱、细胞层数减少，厚度变薄，与正常对照组［（40.73±1.32）μm］相比，hUCMSCs治疗组［（31.28±1.79）μm］与PBS治疗组［（17.21±1.02）μm］及光损伤组［（12.68±1.42）μm］的视网膜外核层厚度均变薄（均为P<0.05）。与光损伤组相比，PBS治疗组和hUCMSCs治疗组视网膜外核层层数显著增加。结论　HUCMSCs移植到光损伤大鼠玻璃体内后能存活、迁移及整合到受损伤视网膜，hUCMSCs玻璃体内移植可抑制光损伤大鼠光感受器的凋亡。     

VP1-Binding Protein Glucose-regulated protein 78 as an important mediator for Enterovirus 71 infecting Human Brain Microvascular Endothelial Cells

Purpose: Enterovirus 71 (EV71) is one of the main pathogens causing hand, foot and mouth disease, which could even induce severe brain damage in some patients. As the underlying mechanism of the invasion and replication process still remains largely unknown, we investigated the role of candidate proteins expressed during EV71 invasion in human brain microvascular endothelial cells (HBMECs) to delineate the pathophysiological mechanism of EV-71 infection. Materials and Methods: Ninety-one candidate EV71-associated proteins which could bind the major capsid protein (viral protein 1 [VP1]) of EV71 on the HBMEC were identified by applying an analysis of glutathione-S-transferase pull-down coupling with liquid chromatography-electrospray ionisation-tandem mass spectrometry (LC-ESI-MS/MS). Seventy-eight kDa glucose-regulated protein 78 (GRP78) binding to the VP1 protein was further validated by co-immunoprecipitation, immunofluorescence and western blot analysis. To explore the role of GRP78 in EV71 infection, GRP78 was knocked down and overexpressed in HBMEC and was verified by TCID50 assay. Results: LC-ESI-MS/MS-identified 91 proteins were subjected to gene ontology analysis, and on molecular and biological function analysis revealed GRP78 act as an important binding protein in mediating EV71 infection. In addition, immunofluorescence demonstrated the co-localisation of GRP78 and VP1 in cytoplasm of the infected HBMEC. The TCID50 assay showed that knockdown of GRP78 could attenuate the replication capacity of EV71 in HBMEC, and the overexpression could increase the virus titre in HBEMC at 24 h post-infection suggesting that GRP78 was associated with the replication capacity of EV71 in HBMEC. Conclusion: These findings provided evidence that GRP78 plays an important role during the progression of EV71 infection as a mediator in HBMEC.     

Effects of Pistacia atlantica on Oxidative Stress Markers and Antioxidant Enzymes Expression in Diabetic Rats

Introduction: Diabetes mellitus (DM) affects many patients all over the world. It involves different parts of the body, such as brain, eyes, kidneys, vessels, and so on. The lack of balance between free radicals and antioxidants is a possible mechanism involved in the pathogenesis of diabetes. Antioxidant treatment, especially natural forms, can be a beneficial solution. Therefore, we evaluated the effects of Pistacia atlantica oleoresin (PAO) on oxidative stress markers and antioxidant enzymes expression in diabetic rats.

Method: Fifty adult male Wistar rats were allotted randomly into five groups as follow: control group, diabetic control group, glibenclamide control group, diabetic glibenclamide group, diabetic treated group with 200?mg/kg PAO. Then PAO was prepared and analyzed by gas chromatography/mass spectroscopy (GC/MS). LD50 was also estimated for essential oil. Oxidative stress markers and antioxidant enzyme including malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GPx), catalase (CAT), and superoxide dismutase (SOD) were also measured. The expression of GPx, CAT, and SOD genes was investigated using real-time polymerase chain reaction (PCR).

Results: The main constituents of essential oil gum were beta-pinene (29.38%), followed by alpha-pinene (18.15%), myrcene (7.36%), trans-pinocarveol (7.15%), and camphene (4.12%). Diabetes induced an increased level of MDA (69.92?±?3.92 vs. 43.76?±?3.73) and decreased levels of GSH (2.57?±?0.40 vs. 7.05?±?1.59), GPx (11.66?±?2.2 vs. 16.38?±?2.1), CAT (12.17?±?3.38 vs. 18.7?±?2.66), and SOD (0.78?±?0.67 vs. 2.41?±?0.46). In contrast, PAO treatment significantly decreased MDA (54.59?±?12.54 vs. 69.92?±?3.92) and increased GSH (4.5?±?0.89 vs. 2.57?±?0.40), GPx (25.86?±?5.37 vs. 11.66?±?2.2), CAT (22.69?±?0.36 vs. 12.17?±?3.38), and SOD (3.65?±?1.08 vs. 0.78?±?0.67) (p?<?0.05). Moreover, our results indicated that both GPx and CAT mRNA levels significantly increased approximately 4.46 and 6.23 times in rats fed with 200?mg/kg of PAO, more than that of the healthy control group, respectively (p?<?0.01 and p?<?0.001, respectively). Also, the average expression level of SOD was also significantly 1.57 higher in rats fed with 200?mg/kg of PAO in comparison to the diabetic control group (p?<?0.05).

Conclusion: The results indicated that PAO could be propose as an agent that protects the body against diseases that are associated with oxidative stress.     

Modelling human pathology of traumatic brain injury in animal models

Traumatic brain injury (TBI) is caused by a head impact with a force exceeding regular exposure from normal body movement which the brain normally can accommodate. People affected include, but are not restricted to, sport athletes in American football, ice hockey, boxing as well as military personnel. Both single and repetitive exposures may affect the brain acutely and can lead to chronic neurodegenerative changes including chronic traumatic encephalopathy associated with the development of dementia. The changes in the brain following TBI include neuroinflammation, white matter lesions, and axonal damage as well as hyperphosphorylation and aggregation of tau protein. Even though the human brain gross anatomy is different from rodents implicating different energy transfer upon impact, especially rotational forces, animal models of TBI are important tools to investigate the changes that occur upon TBI at molecular and cellular levels. Importantly, such models may help to increase the knowledge of how the pathologies develop, including the spreading of tau pathologies, and how to diagnose the severity of the TBI in the clinic. In addition, animal models are helpful in the development of novel biomarkers and can also be used to test potential disease‐modifying compounds in a preclinical setting.