血管紧张素转化酶2基因转染对实验性动脉硬化兔斑块炎症的影响

目的 探讨血管紧张素转化酶2(ACE2)基因转染是否能通过使血管紧张素(Ans)Ⅱ转化为Ang1-7而抑制动脉硬化斑块的炎症反应.方法 克隆小鼠ACE2基因,并构建复制缺陷重组腺病毒质粒Ad-ACE2;用球囊损伤内皮细胞及高脂饲养建立兔动脉硬化模型,喂养3个月,将38只新西兰大白兔随机分为Ad-ACE2及Ad-EGFP两组,每组19只.Ad-ACE2组注射ACE2的腺病毒(2.5×109pfu/m1)入兔的腹主动脉中,Ad-EGFP组注射Ad-EGFP,注射1个月后处死动物,取腹主动脉,检测巨噬细胞和单核细胞趋化因子(MCP-1)蛋白的表达;同时应用实时定量PCR检测MCP-1基因的表达.结果 Ad-ACE2组的动脉硬化斑块中的巨噬细胞阳性表达率(13.6%±4.2%)明显低于Ad-EGFP组(23.6%±6.9%,P<0.01);而基因转染后MCP-1蛋白的表达(13.2%±0.4%)明显低于Ad-EGFP组(25.0%±7.4%,P<0.01).结论 ACE2基因转染抑制了MCP-1的蛋白表达及巨噬细胞浸润程度,提示ACE2基因具有抑制动脉粥样硬化斑块炎症的作用.     

血管紧张素转换酶2过度表达增加兔动脉粥样硬化模型斑块稳定性

该研究是要验证这样的假设：即血管紧张素转换酶2（ACE2）过度表达，通过拮抗ACE的活性和促进血管紧张素Ⅱ转换为血管紧张素1～7，可以提高动脉粥样硬化斑块的稳定性。方法和结果：兔114只通过损伤血管内皮细胞并给予动脉粥样硬化饮食来制作腹主动脉粥样硬化斑块。A组基因治疗4周，B组基因治疗12周，每组再随机分为3个亚组，分别接受重组ACE2表达载体（AdACE2），AdEGFP对照载体和AdACE2＋A779（一种血管紧张素1～7受体拮抗剂）的治疗。     

RNA干扰LOX-1表达对氧化型低密度脂蛋白诱导人脐静脉内皮细胞损伤的保护机制

目的 构建血凝素样氧化型低密度脂蛋白受体1（LOX-1）基因的RNA干扰慢病毒载体并转染人脐静脉内皮细胞。观察干扰LOX-1表达后对氧化型低密度脂蛋白（ox-LDL）诱导内皮细胞损伤的保护机制。方法 针对已经筛选确定的小干扰RNA有效干扰序列,合成慢病毒LV-si-OLR1最佳干扰序列,测定滴度。转染人脐静脉内皮细胞96 h后,通过荧光显微镜观察转染效率,逆转录聚合酶链反应和Western blot检测LOX-1的抑制效率。转染72 h后加入150 mg/L ox-LDL处理24 h;噻唑蓝比色法检测细胞存活率;硝酸还原酶法检测各组内皮细胞培养液一氧化氮（NO）的含量;Western blot检测各组细胞间黏附分子1（ICAM-1）、单核细胞趋化蛋白1（MCP-1）、RhoA、Rho激酶1（ROCK1）、Rho激酶2（ROCK2）的表达。结果 测序结果证实,靶向人LOX-1的干扰慢病毒载体构建成功,包装后慢病毒滴度为6×10¹¹ TU/L。转染组与未转染组比较,LOX-1 mRNA与蛋白的表达明显减少（P<0.01）。与正常对照组相比,ox-LDL处理组能明显减低内皮细胞的存活率及NO的生成量,增高ICAM-1、MCP-1、RhoA、ROCK1、ROCK2的表达（P<0.05）;与ox-LDL处理组相比,干扰LOX-1表达后对ox-LDL诱导的内皮细胞存活率、NO生成量减低和ICAM-1、MCP-1、RhoA、ROCK1、ROCK2表达增高均有明显抑制作用（P<0.05）。结论 干扰LOX-1表达,对ox-LDL诱导内皮细胞引起的ICAM-1、MCP-1高表达和RhoA/Rho激酶信号通路的激活及细胞存活率、NO生成量的减低均有明显抑制作用,起到对内皮细胞损伤的保护作用。本研究为LOX-1作为靶基因治疗动脉粥样硬化提供了实验依据。     

3种中药复方血清对脂多糖诱导的人脐静脉内皮细胞炎症因子

目的　研究四君子汤、血府逐瘀汤、补阳还五汤等3种中药复方血清对脂多糖诱导的人脐静脉内皮细胞（HUVEC）血凝素样氧化低密度脂蛋白受体1（LOX-1）、肿瘤坏死因子α（TNF-α）、血管细胞黏附分子1（VCAM-1）及细胞间黏附分子1（ICAM-1）表达的影响,探讨相关中药复方防治动脉粥样硬化的机制。方法　应用药物血清学方法制备四君子汤、血府逐瘀汤、补阳还五汤含药血清及正常空白血清,以供细胞实验使用。体外培养HUVEC,随机分为6组,即空白对照组、模型组、阿托伐他汀组、四君子汤组、血府逐瘀汤组、补阳还五汤组,用脂多糖刺激2　h后,分别加入阿托伐他汀和中药复方血清干预,24　h后收集细胞,用荧光定量PCR和Western　blot测定LOX-1、TNF-α、VCAM-1、ICAM-1的mRNA和蛋白表达。结果　用脂多糖刺激HUVEC后,引起LOX-1、TNF-α、VCAM-1、ICAM-1的mRNA和蛋白表达显著升高（P<0.01）,分别以血府逐瘀汤、补阳还五汤含药血清及阿托伐他汀干预后可显著抑制LOX-1、TNF-α、VCAM-1、ICAM-1的mRNA和蛋白表达（P<0.05）,而四君子汤含药血清则未见显著影响（P>0.05）。结论　血府逐瘀汤、补阳还五汤可抑制LOX-1、TNF-α、VCAM-1及ICAM-1等炎症因子的表达,这可能是这两种中药复方防治动脉粥样硬化作用的机制之一。而四君子汤未见此作用。     

雷米普利与动脉粥样硬化斑块稳定性关系的实验研究

目的探讨雷米普利与动脉粥样硬化斑块稳定性的关系及机制。方法将24只雄性新西兰大白兔随机分为正常饮食组、高脂饮食组及高脂饮食加雷米普利干预组，应用免疫组化测定兔胸主动脉中植物血凝亲样氧化低密度脂蛋白受体-1（LOX-1）的蛋白表达水平及巨噬细胞在动脉粥样硬化斑块中的浸润水平。用RT-PCR测定兔胸主动脉中LOX-1基因表达的水平。结果高脂组及雷米普利干预组胸主动脉中LOX-1蛋白及基因表达水平明显高于正常饮食组（P〈0．05）；高脂组及雷米普利干预组胸主动脉巨噬细胞在动脉粥样硬化斑块中的浸润水平明显高于正常饮食组；雷米普利干预组胸主动脉中LOX-1蛋白及基因表达水平明显低于高脂饮食组（P〈0．05）；雷米普利干预组胸主动脉巨噬细胞在动脉硬化斑块中的浸润水平明显低于高脂饮食组。结论雷米普利能够明最抑制动脉硬化兔LOX-1蛋白及基因表达，同时明显减少巨噬细胞在动脉硬化斑块中的浸润，对动脉粥样硬化斑块具有稳定作用。     

RNA干扰人血凝素样氧化型低密度脂蛋白受体表达对内皮细胞骨架损伤的影响

目的构建针对人血凝素样氧化型低密度脂蛋白受体1(LOX-1)基因的RNA干扰慢病毒载体,并转染人脐静脉内皮细胞后,采取氧化型低密度脂蛋白(ox-LDL)诱导观察对内皮细胞骨架损伤的影响。方法体外培养人脐静脉内皮细胞分为对照组、ox-LDL组(ox-LDL处理)、阴性转染组(ox-LDL处理+阴性慢病毒转染)和慢病毒转染组(ox-LDL处理+最佳干扰序列慢病毒转染)。荧光显微镜观察转染效率,Western blot检测磷酸化肌球蛋白轻链(p-MLC)、Rho激酶(ROCK)、LOX-1蛋白的表达;免疫荧光法观察细胞骨架肌动蛋白(F-actin)的变化。结果与对照组比较,ox-LDL组和阴性转染组p-MLC、ROCK、LOX-1蛋白表达增高;细胞F-actin发生损伤并伴含量减少(P<0.05)。与阴性转染组比较,慢病毒转染组抑制p-MLC、ROCK、LOX-1蛋白表达,减轻F-actin损伤及伴含量增加(P<0.05)。结论干扰LOX-1表达对ox-LDL诱导引起的内皮细胞ROCK、p-MLC表达增加及细胞骨架损伤均有抑制作用。     

凝集素样氧化型低密度脂蛋白受体-1与动脉粥样硬化关系的研究进展

凝集素样氧化型低密度脂蛋白受体-1（lectin-like oxidized low-density lipoprotein receptor-1,LOX-1）是内皮细胞上氧化型低密度脂蛋白特异性受体,可介导内皮细胞活化、损伤和凋亡,平滑肌细胞和巨噬细胞内脂质聚集,活化血小板促进血栓形成,并增加斑块不稳定性,促进动脉粥样硬化（atherosclerosis,AS）的发生发展。而抑制LOX-1表达可减弱AS病变,提示LOX-1可能成为防治AS的分子靶点。本文就LOX-1与AS的关系作一综述。     

伊贝沙坦对血管紧张素Ⅱ诱导血管内皮细胞NF-κB激活与ICAM-1表达的影响

目的：观察血管紧张素Ⅱ1型受体（AT1R）拮抗剂伊贝沙坦（irbesartan,Irb）对血管紧张素Ⅱ（AngⅡ）诱导的体外培养人脐静脉血管内皮细胞（HUVEC）核因子-κB（NF-κB）激活与细胞问粘附分子-1（ICAM-1）表达的影响。方法体外培养的第3～5代HUVEC用于实验。采用免疫组织化学分析检测细胞NF—κB弧单位p65、ICAM—1的表达程度。结果AngⅡ有效激活NF—κB并诱导HUVEC表达ICAM-1增加Irb预先孵胄2h能抑制NF—κB并减轻AngⅡ诱导的HUVEC ICAM-1表达。结论AngⅡ通过激活NF-κB使ICAM-1表达增加损伤血管内皮功能Irb能抑制NF-κB激活降低HUVEC ICAM-1表达,从而保护血管内皮功能,这有可能减轻心血管疾病损伤的进展。     

SIRT6延缓动脉粥样硬化病程进展的机制研究

目的 分析SIRT6延缓动脉粥样硬化病程进展的机制。方法 本实验时间为2022年1—10月。动物实验：将10只SPF级野生型雄性C57BL小鼠作为对照组，随机选取10只雄性ApoE-/-小鼠作为动脉粥样硬化组，剩余10只雄性ApoE-/-小鼠作为动脉粥样硬化+SIRT6-/-组。对照组小鼠采用普通饲料喂养16周。动脉粥样硬化组和动脉粥样硬化+SIRT6-/-组小鼠采用高脂饲料喂养16周以构建动脉粥样硬化模型。此外，动脉粥样硬化+SIRT6-/-组小鼠采用CreLoxP方法构建巨噬细胞特异性SIRT6缺陷模型。采用HE染色检测各组小鼠动脉粥样硬化斑块面积，Western blot法检测各组小鼠主动脉组织中SIRT6、IL-32、细胞间黏附分子1(ICAM-1)表达水平，免疫荧光染色检测各组小鼠主动脉组织中CD31表达情况。细胞实验：取对数生长期的巨噬细胞RAW 264.7，将其随机分为对照组（不进行干预）、动脉粥样硬化组[采用50μg/ml的氧化型低密度脂蛋白（ox-LDL）干预24 h]、动脉粥样硬化+Ad-SIRT6组（转染腺病毒Ad-SIRT6,24 h后用50μg/ml的ox-L...     

缬沙坦抑制非对称二甲基精氨酸诱导的内皮细胞LOX-1的表达

目的 研究缬沙坦对非对称二甲基精氨酸(ADMA)诱导的人脐静脉内皮细胞(HUVEC)凝集素样氧化型低密度脂蛋白受体1 (LOX-1) mRNA和蛋白表达的影响,并探讨其机制.方法 体外培养HUVEC,以不同浓度的缬沙坦和15 μmol/L ADMA共同孵育24h,DCFH-DA检测细胞内活性氧(ROS)生成量；RT-PCR测定NADPH氧化酶p22phox及LOX-1 mRNA的转录水平；酶联免疫吸附法检测细胞LOX-1蛋白的表达水平.结果 与ADMA组相比,缬沙坦干预后细胞ROS水平显著下降(P＜0.05),p22phox和LOX-1 mRNA转录水平及LOX-1蛋白的表达水平明显下调(P＜0.05),并呈剂量依赖性.结论 缬沙坦通过抑制p22phox的表达减少细胞内ROS水平,进而抑制ADMA诱导的内皮细胞LOX-1 mRNA转录和蛋白表达,这可能是其独立于降压作用的抗动脉粥样硬化作用的部分机制.     

Three layer functional model and energy exchange concept of aging process

Relying on a certain degree of abstraction, we can propose that no particular distinction exists between animate or living matter and inanimate matter. While focusing attention on some specifics, the dividing line between the two can be drawn. The most apparent distinction is in the level of structural and functional organization with the dissimilar streams of ‘energy flow’ between the observed entity and the surrounding environment. In essence, living matter is created from inanimate matter which is organized to contain internal intense energy processes and maintain lower intensity energy exchange processes with the environment. Taking internal and external energy processes into account, we contend in this paper that living matter can be referred to as matter of dissipative structure, with this structure assumed to be a common quality of all living creatures and living matter in general. Interruption of internal energy conversion processes and terminating the controlled energy exchange with the environment leads to degeneration of dissipative structure and reduction of the same to inanimate matter, (gas, liquid and/or solid inanimate substances), and ultimately what can be called ‘death.’ This concept of what we call dissipative nature can be extended from living organisms to social groups of animals, to mankind. An analogy based on the organization of matter provides a basis for a functional model of living entities. The models relies on the parallels among the three central structures of any cell (nucleus, cytoplasm and outer membrane) and the human body (central organs, body fluids along with the connective tissues, and external skin integument). This three-part structural organization may be observed almost universally in nature. It can be observed from the atomic structure to the planetary and intergalactic organizations. This similarity is corroborated by the membrane theory applied to living organisms. According to the energy nature of living matter and the proposed functional model, the decreased integrity of a human body's external envelope membrane is a first cause of the structural degradation and aging of the entire organism. The aging process than progresses externally to internally, as in single cell organisms, suggesting that much of the efforts towards the restoration and maintenance of the mechanisms responsible for structural development should be focused accordingly, on the membrane, i.e., the skin. Numerous reports indicate that all parts of the human body, like: bones, blood with blood vessels, muscles, skin, and so on, have some ability for restoration. Therefore, actual revival of not only aging tissue of the human body's membrane, but the entire human body enclosed within, with all internal organs, might be expected. We assess several aging theories within the context of our model and provide suggestions on how to activate the body's own anti-aging mechanisms and increase longevity. This paper presents some analogies and some distinctions that exist between the living dissipative structure matter and inanimate matter, discusses the aging process and proposes certain aging reversal solutions.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Unusual responses of nocturnal pineal melatonin synthesis and secretion to swimming: Attempts to define mechanisms

Abstract: The effect of swimming at night on rat pineal melatonin synthesis was compared with that of light exposure at night. Rats were forced to swim at 0030 hr (lights out at 2000 hr) and sacrificed by decapitation 15 and 30 min later, immediately after swimming. Other groups of animals were exposed to white light (650μW/cm²) for 15 and 30 min at same time. Swimming caused a rapid and highly significant drop in the melatonin content in the pineal gland; however, the activity of N-acetyltransferase (NAT), the supposed rate limiting enzyme in the melatonin production, was not changed. Despite the drop in pineal melatonin levels, serum concentrations of the indole remained elevated in the rats that swam. In contrast, melatonin levels in the pineal and serum of light exposed rats fell precipitously, accompanied by a significant suppression of NAT activity. Since we anticipated that the strenuous exercise associated with swimming may induce release of artrial natriuretic peptide (ANP) from the heart, which in turn could cause the release of pineal melatonin, in a second study we injected physiological saline intravenously to stretch the cardiac muscle and release ANP. Three milliliters of normal saline was injected during the day into the jugular vein of anesthetized rats that were pretreated with isoproterenol to stimulate pineal melatonin production. Animals were killed 15 min after the saline injection, and pineal NAT activity and pineal melatonin levels were measured. The saline injections caused no alteration in the elevated levels of either NAT or melatonin. These data suggest that the disparity in pineal NAT activity (which was high) and pineal melatonin (which was low), in animals swum at night, may not be caused by ANP which is released during strenuous exercise such as swimming.     

Melatonin and photoperiodic time measurement: Seasonal breeding in the sheep

Abstract: Well-established circadian physiology supports the view that photoperiodic time measurement utilizes the coincidence between the presence of light and a photosensitive phase of a 'biological clock' to alter reproductive status—the so-called external coincidence model of seasonal breeding. In this review, we examine the mechanism whereby photoperiod interacts with presumed suprachiasmatic nuclei activity to allow endogenous melatonin to normally synchronize reproductive activity to the optimal time of year. The Romney Marsh sheep is particularly explored as an experimental model. It is suggested that the on/off activity of seasonal reproduction may be a robust mechanism able to be predictably manipulated by the judicious use of the light/dark cycle and exogenous melatonin, but firmly based on circadian principles.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Language of CTO interventions – Focus on hardware

The knowledge of variety of chronic total occlusion (CTO) hardware and the ability to use them represents the key to success of any CTO interventions. However, the multiplicity of CTO hardware and their physical character and the terminology used by experts create confusion in the mind of an average interventional cardiologist, particularly a beginner in this field. This knowledge is available but is scattered. We aim to classify and compare the currently used devices based on their properties focusing on how physical character of each device can be utilized in a specific situation, thus clarifying and simplifying the technical discourse.