丹参酮生物合成相关SmERF108转录因子的鉴定及其靶基因分析＊

目的 鉴定与丹参酮合成相关转录因子AP2/ERF家族中SmERF108转录因子，并分析转录因子SmERF108的靶基因。方法 利用生物信息学在线分析平台如NCBI、PFAM等分析SmERF108的序列特征；MEGA-X软件用于构建SmERF108与不同功能转录因子的系统进化树；拟南芥原生质体转化法鉴定SmERF108蛋白的亚细胞定位；通过实时荧光定量PCR（Real-time qPCR）对SmERF108基因在丹参不同器官和组织差异表达进行检测；利用酵母体系对SmERF108的转录激活活性进行探究并用酵母单杂技术确定其靶基因。结果 SmERF108具有典型的AP2/ERF保守结构域，属于ERF-B3亚组，系统进化树和保守基序分析显示SmERF108与丹参中SmERF128、青蒿中AaERF2亲缘关系较近且保守基序分布一致；亚细胞定位显示SmERF108蛋白定位于细胞核；SmERF108基因在周皮中表达最高，且呈现周皮（R1）>韧皮部（R2）>木质部（R3）的规律；酵母自激活验证SmERF108具有转录激活活性，同时酵母单杂交确认其能与关键酶基因SmCPS1启动子结合。结论 鉴定到丹参中一个新转录因子SmERF108，生物信息学分析及差异表达分析预测与丹参酮合成相关，分子互作初步证实靶基因为SmCPS1二萜环化酶。     

丹参营养器官中脂溶性成分的动态变化规律

目的 测定丹参营养器官中丹参酮Ⅰ和丹参酮Ⅱ_A的含量,并揭示丹参生长过程中,丹参酮Ⅰ和丹参酮Ⅱ_A的动态变化规律.方法 采用反相高效液相色谱法,以Zorbax SB-C_(18)(150mm×4.6mm, 5.0μm)为色谱柱;甲醇-0.4%磷酸为流动相,梯度洗脱,流速为1.0 ml/min;二极管阵列检测器:检测波长分别为246, 270 nm;柱温为30℃.结果 丹参叶和茎中均未检出丹参酮Ⅰ和丹参酮Ⅱ_A,根中丹参酮Ⅱ_A和丹参酮Ⅰ的含量变化呈"单峰"曲线,丹参酮Ⅱ_A的含量在7月份最高,为2.94%,丹参酮Ⅰ含量在5月份最高,为0.78%.结论 4月至7月是丹参根中丹参酮积累的关键时期.     

Cellular nonmuscle myosins NMHC‐IIA and NMHC‐IIB and vertebrate heart looping

Flectin, a protein previously described to be expressed in a left‐dominant manner in the embryonic chick heart during looping, is a member of the nonmuscle myosin II (NMHC‐II) protein class. During looping, both NMHC‐IIA and NMHC‐IIB are expressed in the mouse heart on embryonic day 9.5. The patterns of localization of NMHC‐IIB, rather than NMHC‐IIA in the mouse looping heart and in neural crest cells, are equivalent to what we reported previously for flectin. Expression of full‐length human NMHC‐IIA and ‐IIB in 10 T1/2 cells demonstrated that flectin antibody recognizes both isoforms. Electron microscopy revealed that flectin antibody localizes in short cardiomyocyte cell processes extending from the basal layer of the cardiomyocytes into the cardiac jelly. Flectin antibody also recognizes stress fibrils in the cardiac jelly in the mouse and chick heart; while NMHC‐IIB antibody does not. Abnormally looping hearts of the Nodal^{Δ 600} homozygous mouse embryos show decreased NMHC‐IIB expression on both the mRNA and protein levels. These results document the characterization of flectin and extend the importance of NMHC‐II and the cytoskeletal actomyosin complex to the mammalian heart and cardiac looping. Developmental Dynamics 237:3577–3590, 2008. © 2008 Wiley‐Liss, Inc.     

How human neutrophils kill and degrade microbes: an integrated view

Summary:  Neutrophils constitute the dominant cell in the circulation that mediates the earliest innate immune human responses to infection. The morbidity and mortality from infection rise dramatically in patients with quantitative or qualitative neutrophil defects, providing clinical confirmation of the important role of normal neutrophils for human health. Neutrophil-dependent anti-microbial activity against ingested microbes represents the collaboration of multiple agents, including those prefabricated during granulocyte development in the bone marrow and those generated de novo following neutrophil activation. Furthermore, neutrophils cooperate with extracellular agents as well as other immune cells to optimally kill and degrade invading microbes. This brief review focuses attention on two examples of the integrated nature of neutrophil-mediated anti-microbial action within the phagosome. The importance and complexity of myeloperoxidase-mediated events illustrate a collaboration of anti-microbial responses that are endogenous to the neutrophil, whereas the synergy between the phagocyte NADPH (nicotinamide adenine dinucleotide phosphate) oxidase and plasma-derived group IIA phospholipase A₂ exemplifies the collective effects of the neutrophil with an exogenous factor to achieve degradation of ingested staphylococci.     

薄层扫描法测定更年康冲剂中丹参酮ⅡA的含量

目的:测定更年康冲剂中丹参酮ⅡA的含量.方法:采用双波长薄层扫描法.结果:丹参酮ⅡA的点样量为0.50～2.50 μg,与吸收度积分值呈良好的线性关系,平均回收率98.50%,RSD＝1.71%.结论:该方法简便、灵敏、重现性好.     

丹参酮对离体肺的保护作用研究

目的观察丹参酮对离体肺的保护作用。方法24只新西兰免随机平均分为两组，对照组用LPD液行肺动脉灌注和保存，实验组用丹参酮加LPD液以同样方法灌注和保存。18h后观察形态学改变。结果光镜下两组肺泡、支气管及毛细血管结构完整，高倍视野下对照组较实验组肺泡上皮及毛细血管上皮细胞肿胀、肥大，少部分组织可见肺泡内有红细胞渗出、间隔增宽等。结论在LPD液中加入丹参酮，能够对离体肺起到更好的保护作用。     

丹参酮诱导HL—60细胞分化及凋亡的流式细胞术分析

应用体外细胞培养及流式细胞仪技术，分析了丹参酮作用前后HL－60细胞的细胞周期分布、细胞增殖指数、p53、c－myc、c－fos及bc1－2基因产物的表达变化，并以全反式维甲酸为对照。结果表明：0．5μg．ml－1的丹参酮作用后，HL－60细胞趋向粒细胞系统分化，有部分细胞发生凋亡（11．8％），丹参酮通过阻止HL－60细胞进入S期而抑制其DNA合成，其作用的分子机制与c－fos基因的表达增高、c－myc基因及bc1－2基因的表达降低有关，诱导分化与细胞凋亡的关系尚有待进一步探讨。     

丹参酮ⅡA磺酸钠注射液对冠心病心肌缺血的治疗作用

目的观察丹参酮ⅡA磺酸钠对冠心病心肌缺血的疗效及安全性。方法将77例冠心病患者随机分为A组40例与B组37例。B组常规应用硝酸异山梨酯与阿司匹林。A组在常规治疗基础上加用丹参酮ⅡA碘酸钠注射液。每例于治疗开始前1 d与结束次日,进行血脂、血液流变学与动态心电图(Holter)检测。结果A组查血脂示血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)明显下降(P均<0.01),B组TC、TG、LDL-C治疗前后均无显著性差异(P均>0.05)。A组血液流变学观察示低切变率下全血黏度、高切变率下全血黏度、血浆比黏度、红细胞聚集指数、红细胞比积、血小板聚集率、纤维蛋白原均显著下降(P<0.01或0.05),其改善血液流变学指标的疗效优于B组(P<0.05)。Holter监测示症状性心肌缺血与无症状心肌缺血的发作次数及其持续时间2组均明显减少(P<0.01或0.05),但A组比B组减少更为显著(P均<0.05)。在随访治疗过程中未见丹参酮ⅡA磺酸钠的严重不良反应。结论丹参酮ⅡA磺酸钠有降脂与改善血液流变性作用,是治疗冠心病心肌缺血的一种有效而安全的药物。     

甘西鼠尾草中丹参酮IIA的含量测定

目的测定甘西鼠尾草中丹参酮IIA的含量。方法采用HPLC法,色谱柱:ZORB-AX Eclipe XDB-C18(250×4.6mm,5μm)柱,流动相为:甲醇-水(75:25v/v),检测波长:270nm。结果丹参酮IIA在6.0~100.0μg.m l-1范围内,峰面积与进样浓度线性关系良好,回归方程Y=186.3 245.1C,r=0.9974,平均回收率为98.0%,RSD=1.67%;栽培品甘西鼠尾草中丹参酮IIA含量:1年生0.305%,2年生0.539%,3年生0.776%,野生品0.573%。结论所测样品中丹参酮IIA含量均高于《中国药典》2005年版丹参项下规定要求,且栽培3年生甘西鼠尾草其丹参酮IIA含量高于野生品。     

Tanshinone IIA protects rat primary hepatocytes against carbon tetrachloride toxicity via inhibiting mitochondria permeability transition

Tanshinone IIA (Tan IIA), one of the key components of Salvia milthorrhiza Bunge (Lamiaceae), is used to treat liver disease. The present study was carried out to investigate the possible mechanisms involved in the hepatoprotective effects of Tan IIA on carbon tetrachloride (CCl₄)-induced hepatocyte toxicity. In cultures treated with 1 or 2 μM CCl₄, Tan IIA (10–75 μM) significantly increased hepatocyte survival rates. However, only at a concentration of 75 μM could Tan IIA partially reverse the CCl₄ (3 μM)-induced decrease of survival rate (34?±?3% vs. 18?±?3%, n?=?8, p?<?0.01). In isolated mitochondria energized with succinate, Tan IIA could inhibit the large swelling effect induced by CCl₄ (1 and 2 μM). Base on these results, Tan IIA could protect rat primary cultured hepatocytes from CCl₄-induced toxicity partially by the inhibitory effect on the opening of mitochondrial permeability transition (MPT).