玉米苞叶对动脉粥样硬化家兔平滑肌细胞凋亡及p53和Fas蛋白表达的影响

目的：探讨玉米苞叶防治动脉粥样硬化（AS）的机制。方法：选用大耳白家兔，复制家兔AS模型，随机分为动脉粥样硬化模型组，玉米苞叶组和正常对照组，成模后给予玉米苞叶煎剂治疗，8w后处死家兔，采用流氏细胞术检测平滑肌细胞的凋亡率以及凋亡相关基因p53和Fas蛋白表达。结果：模型组血管平滑肌细胞的凋亡率明显高于对照组（P＜0．05），p53和Fas蛋白的表达增强（P＜0．05），主动脉壁肉眼观测出现典型斑块。玉米苞叶组平滑肌细胞的凋亡率明显低于模型组（P＜0．05），p53和Fas蛋白表达下调（P＜0．05）；主动脉斑块面积较模型组明显减小，结论：玉米苞叶通过调节p53和Fas蛋白表达而调节AS家兔平滑肌细胞凋亡。     

木贼提取物对大鼠动脉粥样硬化早期血管平滑肌细胞增殖与凋亡的影响

目的研究木贼正丁醇提取物对大鼠动脉粥样硬化（AS）早期主动脉平滑肌细胞增殖及凋亡的影响。方法选用雄性SD大鼠，随机分为正常对照、模型、阻性药对照及提取物组。复制食饵性AS早期模型，同时给予提取物预防性给药。以吉非罗齐为阳性药对照。实验9w，观察主动脉壁病理形态学变化，流式细胞术定量检测主动脉壁平滑肌增殖及凋亡率。结果木贼正丁醇提取物组平滑肌凋亡率明显高于模型组（P〈0．01），增殖指数低于模型组（P〈0．01）。结论木贼正丁醇提取物可促进AS早期平滑肌凋亡，抑制其增殖，具有阻断AS早期病变进展的作用。     

玉米苞叶对动脉粥样硬化家兔血管平滑肌细胞凋亡及Bcl-2和p53蛋白表达的影响

目的探讨玉米苞叶煎剂对实验性动脉粥样硬化(AS)家兔血管平滑肌细胞(VSMCs)凋亡及凋亡相关基因Bcl-2和p53蛋白表达的影响。方法选用大耳白家兔,利用高脂饲料复制家兔动AS模型,随机分为模型组、玉米苞叶煎剂组和正常对照组。实验9 w后确认两组动物造模成功,其中一组给予玉米苞叶煎剂治疗,28 m l/kg.d-1(玉米苞叶1.4 g/m l),实验17 w后,处死家兔,观察主动脉内膜的病理形态学改变,采用流式细胞术检测VSMCs凋亡率以及凋亡相关基因Bcl-2和p53蛋白表达。结果模型组VSMCs凋亡率明显高于对照组(P<0.05),p53和Bcl-2蛋白表达均增强(P<0.05),肉眼观察,主动脉内膜出现典型的AS斑块,动脉管腔极度狭窄。玉米苞叶煎剂组VSMCs凋亡率明显降低(P<0.05);p53和Bcl-2蛋白表达均下调(P<0.05);主动脉斑块面积较模型组明显减小,动脉管腔狭窄减轻。结论玉米苞叶通过影响p53和Bcl-2蛋白表达而影响AS家兔VSMCs凋亡;同时缩小AS斑块面积,具有显著治疗家兔AS作用。     

玉米苞叶降低动脉硬化家兔白细胞凋亡及CD44表达

目的　探讨玉米苞叶对动脉粥样硬化模型动物外周血白细胞凋亡和CD4 4表达的影响。方法　将家兔分为正常对照组、动脉硬化模型组、低剂量组 (造模 9w后给予低剂量玉米苞叶煎剂 )、高剂量组 (造模 9w后给予高剂量玉米苞叶煎剂 )。 1 7w后测血脂和血中白细胞凋亡率及CD4 4表达量 ,观察主动脉形态变化。结果　①模型组白细胞凋亡率 (6 58± 2 53) % ,CD4 4表达量 6 97± 1 41 ,均显著高于正常组 (P <0 0 1 ) ,②白细胞凋亡率和CD4 4表达量在低剂量组分别为 3 1 1± 1 50、4 76± 0 57;高剂量组为 3 74± 1 66、4 64± 0 42 ,均显著低于模型组 (P <0 0 1 )。结论　玉米苞叶煎剂可显著降低动脉硬化家兔白细胞凋亡率和CD4 4表达量 ,且这种作用不是通过降血脂实现的。     

玉米苞叶煎剂对动脉粥样硬化家兔血管外器官病变的形态学影响

目的 观察玉米苞叶煎剂对AS家兔血管外各器官病变的形态学影响.方法 选用大耳白家兔,雌雄各半,随机分为对照组、高脂组、玉米苞叶煎剂组,复制高脂食饵性家兔AS模型,观察玉米苞叶煎剂预防性用药后对家兔血脂及内脏各器官的病理形态学影响.结果 玉米苞叶煎剂组TC、TG较高脂组明显降低(P<0.01);高脂组各脏器(心、肝、脾、肺、肾)与对照组相比均有明显病理变化,玉米苞叶煎剂组各脏器病变较高脂组明显减轻.结论 玉米苞叶煎剂不但可以减轻AS家兔血管病变,且对心、肝、脾、肺、肾各脏器的高脂损伤同样有干预作用.     

木贼正丁醇萃取物对高脂血症大鼠血管内皮黏附分子表达的影响

目的研究木贼正丁醇萃取物对高脂血症大鼠血脂及主动脉内皮细胞黏附功能的影响。方法选用雄性SD大鼠,随机分为正常对照组、病理模型组、吉非罗齐组及木贼正丁醇治疗组。复制食饵性动脉粥样硬化早期动物模型,用木贼萃取物预防性给药,以吉非罗齐为阳性药对照。实验9 w,检测血脂,流式细胞术定量检测主动脉内皮细胞ICAM-1、VCAM-1的表达及凋亡率,光镜下观察主动脉形态学变化。结果木贼萃取物组血清TG、VLDL水平较模型组显著下降(P<0.05),内皮ICAM-1表达较模型组明显下降(P<0.05),VCAM-1有下降趋势(P>0.05),凋亡率显著下降(P<0.05);形态学显示木贼萃取物组主动脉内皮损伤程度较模型组减轻。结论木贼正丁醇萃取物可保护血管内皮,降低内皮细胞黏附分子表达,阻止动脉粥样硬化早期炎症反应的发生。     

健中愈疡片对乙酸致胃溃疡大鼠胃粘膜细胞动力学的影响

目的研究健中愈疡片对乙酸致大鼠胃溃疡边缘粘膜细胞凋亡和细胞增殖的影响.方法制备乙酸致大鼠胃溃疡模型.分别予健中愈疡片、雷尼替丁和生理盐水治疗14 d后,测量溃疡面积,检测胃溃疡边缘粘膜细胞凋亡指致、增殖细胞核抗原标记指数和Bcl-2蛋白表达.结果健中愈疡片组和雷尼替丁组的溃疡面积明显小于生理盐水组(P＜O.O1).与胃溃疡模型组、雷尼替丁组和生理盐水组比较,健中愈疡片组的溃疡边缘细胞凋亡指数明显下降(P＜O.01),Bcl-2蛋白表达和增殖细胞核抗原标记指数均显著增加(P＜O.01).结论健中愈疡片具有减少胃溃疡边缘粘膜细胞凋亡和促进细胞增殖的作用,这可能是其治疗胃溃疡的作用机制之一.     

两种木贼提取物对动脉粥样硬化早期模型大鼠平滑肌凋亡及BCl-2、Caspase-3表达的调控

目的研究木贼正丁醇提取物及提取剩余物对动脉粥样硬化(AS)早期大鼠主动脉平滑肌细胞凋亡率及Bcl-2、Caspase-3表达的影响。方法雄性SD大鼠随机分为正常对照、模型、阳性药、提取物及提取剩余物组。复制食饵性AS早期模型,用木贼提取物、提取剩余物分别预防性给药,以吉非罗齐为阳性药对照。9w后,观察主动脉壁病理学变化,流式细胞术定量检测主动脉壁平滑肌细胞凋亡率及Bcl-2、Caspase-3的表达。结果木贼正丁醇提取物及提取剩余物组的凋亡率明显高于模型组(P<0.01);Bcl-2低于模型组(P<0.01);Caspase-3高于模型组(P<0.01)。结论木贼正丁醇提取物及提取剩余物通过控制AS早期主动脉壁平滑肌细胞Bcl-2、Caspase-3表达,促进平滑肌细胞凋亡,阻止AS的进程。     

缺氧复氧对平滑肌细胞增殖与凋亡的影响及蝎毒的干预

目的 观察缺氧复氧及在此基础上给予蝎毒后血管平滑肌细胞(VSMC)增殖和凋亡的改变及凋亡相关基因bcl-2、bax的变化,探讨缺氧复氧对VSMC的损伤和蝎毒对缺氧复氧VSMC的保护作用及机制.方法 组织贴块法培养VSMC后随机分为6组:对照组、缺氧复氧模型组、模型+蝎毒组,其中蝎毒又分为5、10、20、40 mg/L不同浓度组.MTT测定细胞增殖变化,TUNEL法检测细胞凋亡,免疫组化法测定bcl-2、bax表达.结果 (1)MTT法测定对照组、模型组A值分别为1.70±0.24、2.23±0.27,二者存在显著性差异(P＜0.05).蝎毒5、20 mg/L组A值分别为1.99±0.28、2.13±0.24,与对照组比较有统计学意义(P＜0.05);10、40 mg/L组A值分别为1.92±0.16、1.89±0.74,与模型组比较有统计学意义(P＜0.05).(2)对照组细胞凋亡率为(26.4±1.4)%,模型组凋亡率为(17.4±0.9)%,两组之间存在显著差异(P＜0.05).蝎毒5、10、40 mg/L组细胞凋亡率分别为(25.6±2.8)%、(33.5±3.5)%、(34.5±5.2)%,与模型组比较均有统计学意义(均P＜0.05);20 mg/L组细胞凋亡率为(20.1±2.4)%,与对照组比较有统计学意义(P＜0.05).(3)与对照组比较,模型组bcl-2与bax蛋白阳性率及bcl-2/bax比值均有显著变化(P＜0.05);蝎毒各浓度组bcl-2/bax比值与模型组比较均降低(P＜0.05).结论 缺氧复氧后VSMC增殖增多,凋亡减少,蝎毒可以抑制缺氧复氧后VSMC的增殖,促进凋亡,并通过凋亡相关基因bcl-2、bax通路,使bcl-2/bax蛋白比值变化而介导细胞凋亡.     

红花对AngⅡ诱导大鼠胸主动脉血管平滑肌细胞凋亡和Caspase-3表达的影响

目的 观察中药红花对血管紧张素Ⅱ(AngⅡ)诱导大鼠动脉平滑肌细胞增殖和Caspase-3表达的影响.方法 培养大鼠胸主动脉血管平滑肌细胞,将经3次传代生长良好的大鼠平滑肌细胞用无血清的DMEM/F-12培养基培养24h,随机分为模型组、红花组、DMSO对照组、空白对照组.模型组、红花组均加AngⅡ、DMSO,同时红花组加入红花,DMSO组仅加入DMSO,空白对照组加入等剂量培养基;采用原位末端标记技术检测凋亡细胞,免疫组化法检测Caspase-3表达.结果 模型组、红花组、DMSO对照组、空白对照组细胞凋亡率分别为4.44％ ±1.81％、19.94％ ±3.92％、3.96％±1.77％、0,红花组与其他三组比较,P均＜0.01,模型组、DMSO对照组与空白对照组比较,P均＜0.05.模型组、红花组、DMSO对照组、空白对照组Caspase-3表达的灰度值分别为134.29±16.86、147.87±12.43、136.20±15.21、130.19±14.32,四组间两两比较,P均＞0.05.结论 红花可以显著促进AngⅡ诱导的大鼠动脉平滑肌细胞凋亡作用,其促进凋亡的途径与Caspase-3蛋白表达无明显关联.     

Role of white cells and neoplastic cells in haemostasis

The aim of this review was to assess the role of white cells and neoplastic cells in haemostasis.     

Metabolic state of glioma stem cells and nontumorigenic cells

Gliomas contain a small number of treatment-resistant glioma stem cells (GSCs), and it is thought that tumor regrowth originates from GSCs, thus rendering GSCs an attractive target for novel treatment approaches. Cancer cells rely more on glycolysis than on oxidative phosphorylation for glucose metabolism, a phenomenon used in 2-[(18)F]fluoro-2-deoxy-D-glucose positron emission tomography imaging of solid cancers, and targeting metabolic pathways in cancer cells has become a topic of considerable interest. However, if GSCs are indeed important for tumor control, knowledge of the metabolic state of GSCs is needed. We hypothesized that the metabolism of GSCs differs from that of their progeny. Using a unique imaging system for GSCs, we assessed the oxygen consumption rate, extracellular acidification rate, intracellular ATP levels, glucose uptake, lactate production, PKM1 and PKM2 expression, radiation sensitivity, and cell cycle duration of GSCs and their progeny in a panel of glioma cell lines. We found GSCs and progenitor cells to be less glycolytic than differentiated glioma cells. GSCs consumed less glucose and produced less lactate while maintaining higher ATP levels than their differentiated progeny. Compared with differentiated cells, GSCs were radioresistant, and this correlated with a higher mitochondrial reserve capacity. Glioma cells expressed both isoforms of pyruvate kinase, and inhibition of either glycolysis or oxidative phosphorylation had minimal effect on energy production in GSCs and progenitor cells. We conclude that GSCs rely mainly on oxidative phosphorylation. However, if challenged, they can use additional metabolic pathways. Therefore, targeting glycolysis in glioma may spare GSCs.     

Ex vivo expansion of hematopoietic progenitor cells and mature cells

Hematopoietic cells have the potential for providing benefit in a variety of clinical settings. These include cells for support of patients undergoing high-dose chemotherapy, as a target for replacement gene therapy, and as a source of cells for immunotherapy. The limitation to many of these applications has been the total absolute number of defined target cells. Therefore many investigators have explored methods to culture hematopoietic cells in vitro to increase the numbers of these cells. Studies attempting to expand hematopoietic stem cells, progenitor cells, and mature cells in vitro have become possible over the past decade due to the availability of recombinant growth factors and cell selection technologies. To date, no studies have demonstrated convincing data on the expansion of true stem cells, and so the focus of this review is the expansion of committed progenitor cells and mature cells. A number of clinical studies have been preformed using a variety of culture conditions, and several studies are currently in progress that explore the use of ex vivo expanded cells. These studies will be discussed in this review. There are evolving data that suggest that there are real clinical benefits associated with the use of the expanded cells; however, we are still at the early stages of understanding how to optimally culture different cell populations. The next decade should determine what culture conditions and what cell populations are needed for a range of clinical applications.     

干细胞、肿瘤干细胞与肿瘤发生

肿瘤干细胞是肿瘤研究的一个新热点,指出肿瘤可能是由肿瘤干细胞产生,肿瘤干细胞则由正常干细胞恶变形成.正常干细胞的特有性状,使其较成体细胞更易成为肿瘤发生的靶细胞.干细胞可能经基因突变、异常不对称分裂和细胞融合转化为肿瘤干细胞.利用不同的蛋白标志物或荧光探针,通过流式细胞仪分选是发现肿瘤干细胞的主要方法.已证实的肿瘤干细胞皆具有强大的自我更新和增殖能力,以及细胞分化潜能.针对肿瘤干细胞的检测和杀伤,可能为肿瘤早期诊断和治疗带来希望.     

胃癌细胞和胃癌多药耐药细胞中P-gp及GST的表达

目的比较胃腺癌SGC-7901细胞和多药耐药细胞SGC-7901/ADR中P-gp和GST表达的差异,进一步探讨胃癌多药耐药机制。方法常规培养人胃癌SGC-7901细胞和多药耐药细胞SGC-7901/ADR。制备细胞爬片,免疫细胞化学方法和灰度测定检测SGC-7901细胞和SGC-7901/ADR中P-gp及GST的表达。结果免疫细胞化学SP法染色后,P-gp蛋白阳性者在胃癌细胞的胞浆和胞膜中可见棕黄色颗粒沉着,GST在胃癌细胞的胞浆中可见棕黄色颗粒。P-gp、GST在胃癌SGC-7901细胞中呈中度表达,在胃癌SGC-7901/ADR细胞中呈高表达,两者比较有显著性差异（P〈0.05）。免疫细胞化学灰度定量测定显示同样的结果。结论 P-gp、GST在胃癌SGC-7901/ADR细胞中的表达较SGC-7901细胞明显增加,可能是胃癌多药耐药的原因之一。     

Differentiation of human embryonic stem cells and human induced pluripotent stem cells into steroid-producing cells

Although there have been reports of the differentiation of mesenchymal stem cells and mouse embryonic stem (ES) cells into steroid-producing cells, the differentiation of human ES/induced pluripotent stem (iPS) cells into steroid-producing cells has not been reported. The purpose of our present study was to establish a method for inducing differentiation of human ES/iPS cells into steroid-producing cells. The first approach we tried was embryoid body formation and further culture on adherent plates. The resultant differentiated cells expressed mRNA encoding the steroidogenic enzymes steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, cytochrome P450-containing enzyme (CYP)-11A1, CYP17A1, and CYP19, and secreted progesterone was detected in the cell medium. However, expression of human chorionic gonadotropin was also detected, suggesting the differentiated cells were trophoblast like. We next tried a multistep approach. As a first step, human ES/iPS cells were induced to differentiate into the mesodermal lineage. After 7 d of differentiation induced by 6-bromoindirubin-3'-oxime (a glycogen synthase kinase-3β inhibitor), the human ES/iPS cells had differentiated into fetal liver kinase-1- and platelet derived growth factor receptor-α-expressing mesodermal lineage cells. As a second step, plasmid DNA encoding steroidogenic factor-1, a master regulator of steroidogenesis, was introduced into these mesodermal cells. The forced expression of steroidogenic factor-1 and subsequent addition of 8-bromoadenosine 3',5'-cyclic monophosphate induced the mesodermal cells to differentiate into the steroidogenic cell lineage, and expression of CYP21A2 and CYP11B1, in addition to steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, CYP11A1, and CYP17A1, was detected. Moreover, secreted cortisol was detected in the medium, but human chorionic gonadotropin was not. These findings indicate that the steroid-producing cells obtained through the described multistep method are not trophoblast like; instead, they exhibit characteristics of adrenal cortical cells.     

Production of interferons by dendritic cells, plasmacytoid cells, natural killer cells, and interferon-producing killer dendritic cells.

The capacity of mouse spleen conventional dendritic cells (cDCs) and plasmacytoid dendritic cells (pDCs) to produce interferon-gamma (IFN-gamma) or IFN-alpha was assessed, and compared with that of natural killer (NK) cells and the recently identified interferon-producing killer dendritic cells (IKDCs), both of which are frequent contaminants in DC preparations. Fully developed cDCs or pDCs, if free of NK cells or IKDCs, showed little capacity for IFN-gamma production. However, an early developmental form of the CD4-8+ cDC subtype, and the Ly6C- Ly49Q- pDC subtype, both were able to produce moderate amounts of IFN-gamma, although less than IKDCs. In response to toll-like receptor 9 stimuli, both the Ly6C+ Ly49Q+ and the Ly6C- Ly49Q- pDC subtypes were effective producers of IFN-alpha. However, IKDCs, which efficiently produced IFN-gamma and showed immediate cytotoxicity on NK target cells, did not produce IFN-alpha under these conditions.     

侧群细胞与肝癌干细胞

肝癌是严重威胁人类生命和健康的一种疾病.其病因和发病机制尚不完全清楚,治疗缺少有效靶点.对肝癌恶性生长、转移及复发机制的研究正在逐渐深入.近年来的研究认为,肿瘤中存在一小群具有自我更新和分化潜能的细胞,即肿瘤干细胞,可能是肿瘤转移和复发的根源.肝癌中应同样存在这样的一群细胞.侧群(side population,SP)细胞是肿瘤细胞中一小部分,具备干细胞的多种特性且易于分离.肝癌组织中SP细胞的鉴定和分离有可能找到肝癌干细胞,有助于肝癌的转移和复发机制的研究,并为肝癌治疗提供有效治疗靶点.     

Natural killer cells and natural killer T cells

NK cells are important in protecting against viral infections, and they may regulate the immune response. They are activated by hematopoietic blasts and pose a barrier to bone marrow transplantation. They are also abundant in the pregnant uterine decidua, although their role there is unknown. NK cells are normally inhibited from responding to host cells by inhibitory receptors that recognize self class I MHC antigens. There is evidence that NK cells may be important in the regulation of autoimmunity, but there is even stronger evidence that NKT cells regulate autoimmunity. The mechanisms by which these cells are activated and by which they regulate other cells are now being understood at the molecular level.