丹参诱导骨髓基质细胞神经元样细胞分化的microRNA-128路径

目的　研究microRNA-128在丹参制剂诱导大鼠骨髓基质细胞(bone marrow strowal cells,BMSCs)向神经元样细胞分化过程中的调控作用。  方法　培养鉴定大鼠BMSCs,应用反义寡居核苷酸转染技术（Lipofectamin 2000）转染microRNA-128 inhibitors于BMSCs;取转染和未转染的BMSCs,待细胞增殖到60%~70%融合时进行预诱导分化,预诱导24 h后,进行诱导分化;根据处理方式不同分为4组：未诱导组（A组）,丹参制剂诱导组（B组）,转染未诱导组（C组）,转染诱导组（D组）。流式细胞术鉴定BMSCs;qPCR检测microRNA-128及神经元特异性烯醇化酶（NSE）、巢蛋白（Nestin）、微管蛋白（β3-tubulin）的mRNA表达;Western blot检测NSE、Nestin和β3-tubulin的蛋白表达。  结果    BMSCs流式细胞检测CD29和CD90双阳性率达99.17%,CD11b和CD45双阴性率达99.21%;丹参制剂诱导及microRNA-128 inhibitors转染后,细胞microRNA-128表达量显著降低（P<0.05）;诱导与未诱导组,转染与未转染组比较,NSE、Nestin和β3-tubulin的mRNA和蛋白表达均显著升高（P<0.05）。  结论　丹参制剂能够抑制BMSCs的microRNA-128表达,进而影响BMSCs神经分化相关蛋白的表达,促进BMSCs向神经元样细胞分化。     

丹参多酚酸盐对持续性＋Gz 环境下大鼠牙髓的影响

目的：探讨丹参多酚酸盐（DSSM）对高正加速度（＋Gz）重复作用下的大鼠磨牙牙髓的作用。方法：27只 SD 雄性大鼠用随机区组法分为9组,每组3只。阳性对照组和药物组分别在＋5 Gz、＋10 Gz 的条件下共离心30 s,每天重复5次,间隔以＋1Gz 持续60 s,4 d /周,共3周。实验期间药物组于每次离心前30 min 按剂量腹腔注射 DSSM溶液（低、中、高给药剂量分别为2、6、18 mg/kg）,阴性及阳性对照组注射等体积生理盐水。实验结束后,光镜下观察各组大鼠牙髓组织形态学变化情况,并采用 Real time qPCR 方法测定大鼠磨牙牙髓中热休克蛋白（HSP70）mRNA 相对含量表达的情况。结果：＋5 Gz 和＋10 Gz阳性对照组大鼠活动程度降低,磨牙牙髓中出现空泡性变,组间差异不明显。HSP70 mRNA 的相对含量比阴性对照组高（P ＜0．05）。药物组中 HSP70 mRNA 的相对表达量较各自阳性对照组降低,＋10 Gz 高剂量组有统计学意义（P ＜0．05）。结论：＋Gz可引起牙髓损伤,＋Gz 值的增加与 HSP70 mRNA 相对表达量增加相关,DSSM可能对牙髓有保护作用。     

Antimicrobial Activity of Some Endemic Verbascum,Salvia, and Stachys Species

Methanol extracts obtained from endemic Stachys sivasica Kit Tan & Yildiz, Stachys anamurensis Sumbul, Stachys cydni Kotschy ex Gemici & Leblebici, Salvia aytachii Vural & Adiguzel, and Verbascum gypsicola Vural & Aydogdu have been investigated for their antimicrobial activity. Antimicrobial activity was determined with Escherichia coli ATCC 11230, Stapylococcus aureus ATCC 6538P, Klebsiella pneumoniae UC57, Pseudomonas aeruginosa ATCC 27853, Proteus vulgaris ATCC 8427, Bacillus cereus ATCC 7064, Mycobacterium smegmatis CCM 2067, Listeria monocytogenes ATCC 15313, Micrococcus luteus CCM 169, Candida albicans ATCC 10231, Rhodotorula rubra DSM 70403, and Kluyveromyces fragilis ATCC 8608 by the disk-diffusion method. Verbascum gypsicola extracts had strong antimicrobial activity against the gram-positive bacteria and the yeast cultures. The extracts of Stachys L. were effective only against bacteria. The extracts of Salvia aytachii demonstrated an antimicrobial effect against bacteria and the yeast cultures used in this study.     

The interaction of ancrod with human platelets

Ancrod, a serine protease purified from the venom of Agkistrodon rhodostoma, has been used as a therapeutic anticoagulant for a number of indications, including replacement of heparin in patients with heparin-induced thrombocytopenia. Ancrod has similar fibrinolytic activity to thrombin, but ancrod specifically cleaves only the alpha chain of fibrinogen, producing the characteristic fibrinopeptides A, AP and AY. Because ancrod has been used in patients with heparin-induced thrombocytopenia, it is important to ensure that ancrod does not directly affect the platelets and potentially increase the hemostatic effect. The effect of ancrod on platelets has not been well established, and there is not agreement in published studies. Additionally, some of the studies are over 15 years old and pre-date sensitive assays such as glycoprotein analysis. For these reasons, we investigated the interaction of ancrod with human platelets using direct and indirect, functional and biochemical techniques. Incubation of platelets with ancrod alone did not induce platelet aggregation or the release of dense-granule contents. Pre-incubation of platelets with ancrod did not augment or inhibit the maximal aggregation achieved with thrombin, nor did it affect the amount of serotonin release from dense granules caused by activation by thrombin. Studies of ancrod-treated platelets using monoclonal antibody-mediated radioimmunoprecipitation demonstrated that high concentrations of ancrod did not cause measurable cleavage of either the glycoproteins Ib-IX or IIb-IIIa. Incubation of radiolabeled platelets with ancrod-treated plasma also had no effect on the platelet glycoproteins, indicating that ancrod does not indirectly affect the major surface receptors. Direct binding studies using radiolabeled ancrod did not demonstrate specific binding to the platelet surface. Together these studies indicate that ancrod does not directly affect nor bind to platelets in vitro . The hypocoagulant state and subsequent platelet function defect resulting from the use of ancrod appears to be limited to the removal of fibrinogen from the circulation.     

丹参对重度烧伤器官缺血-再灌注损伤保护作用的临床观察

[目的]探讨丹参对严重烧伤后重要脏器功能的治疗、促进创面愈合的临床疗效。[方法]选取2006年12月～2010年5月住院的74例重度烧伤病人,采用随机对照方法分为丹参治疗组(37例)和对照组(37例),治疗组应用注射用丹参冻干20mg/kg静滴,观察用药后5d、10d血常规、肝功能、肾功能和心肌酶谱、血清肿瘤坏死因子-α(TNF-α)及白介素-6(IL-6)水平,器官功能不全(OD)的发生率及病死率及平均愈合时间(d),并统计学分析。[结果]d5治疗组与对照组的各项检验指标改善有差异,P﹤0.05;d10部分检验指标改善有明显差异;治疗组5例出现OD,对照组10例死亡1例。[结论]注射用丹参冻干药用有活血化瘀的功效;烧伤早期大剂量使用可明显改善重要脏器的微循环,减轻缺血再灌注损伤,有效支持和保护重要脏器功能,能有效防止OD的发生、发展。     

Stephania tetrandra prevents and regresses liver fibrosis induced by carbon tetrachloride in rats

Background and Aim:  There is currently no safe and effective treatment for liver fibrosis. We have previously shown that Stephania tetrandra ( ST ) and Salvia miltiorrhiza ( SM ) suppress cell proliferation and enhance apoptosis of hepatic stellate cell (HSC) in vitro . In this study, we aimed to investigate the anti-fibrotic effect of these two herbs in vivo .
Methods:  Liver fibrosis was induced by carbon tetrachloride (CCl₄) injection in rats for 5 weeks. SM , ST or SM  +  ST was gavaged on day 1 of CCl₄ administration to study the preventive effects of herbs on hepatic fibrosis. In a separate study designed to assess possible fibrosis regression, rats were randomly allocated to be treated with SM, ST or SM  +  ST when fibrosis was established. Liver injury and collagen content were assessed. HSC activation and apoptosis were determined.
Results:  As compared with the CCL₄-only rats, serum ALT was significantly lower in CCl₄-treated rats that received either SM ( P  < 0.01) or ST ( P  < 0.01). Administration of ST significantly prevented ( P  < 0.01) or reversed the hepatic fibrosis ( P  < 0.01) induced by CCL₄. Moreover, rats treated with ST had reduced protein expression of α-SMA both in prevention ( P  < 0.05) and in regression ( P  < 0.01) experiments. The double-color staining of α-SMA and TUNEL showed that ST increased HSC apoptosis. However, co-treatment of SM  +  ST did not increase the antifibrotic effect of ST .
Conclusions:  Stephania tetrandra safely and effectively prevents and reverses hepatic fibrosis through activating HSC apoptosis in rats.     

丹参注射液诱导大鼠骨髓间充质干细胞分化为神经元样细胞的研究

目的:探讨丹参注射液在体外诱导大鼠骨髓间充质干细胞(MSCs)向神经元样细胞分化的作用。方法:采用差速贴壁法体外分离培养大鼠MSCs,流式细胞仪检测其表面标志,经碱性成纤维细胞生长因子(bFGF)预诱导24h,采用含丹参注射液的无血清L-DMEM培养液诱导MSCs,倒置显微镜下观察细胞形态的变化,免疫荧光细胞化学检测巢蛋白、神经元特异性烯醇化酶(NSE)、神经丝蛋白(NF-M)、胶质纤维酸性蛋白(GFAP)的表达。结果:体外培养的大鼠MSCs表达CD29、CD44、CD105、CD166。丹参注射液诱导后MSCs胞体收缩,突起伸出,较密的部分神经元拉成网状,免疫细胞化学显示巢蛋白、NSE、NF-M表达阳性,阳性率分别为(58.68±4.50)%、(61.23±5.72)%、(63.47±6.38)%,GFAP阳性细胞率(1.47±0.23)%。结论:丹参注射液在体外可以将大鼠MSCs诱导分化为神经元样细胞。     

丹参不同提取方法中水溶性成分含量的比较研究

目的研究不同提取方法提取的丹参药材中水溶性成分的含量。方法采用高效液相色谱法比较了煎煮、超声、索氏提取3种不同提取方法中丹酚酸B、丹参素、原儿茶醛的含量。结果超声提取丹酚酸B含量最高,索氏提取丹参素、原儿茶醛含量最高。结论测定丹酚酸B的含量宜采用超声法提取,而测定丹参素、原儿茶醛的含量则宜采用索氏提取法。     

复方丹参方制剂治疗稳定型心绞痛随机对照试验的系统评价

[目的]评价复方丹参方制剂(与硝酸酯类作对照)治疗稳定型心绞痛的疗效和安全性。[方法]通过检索Cochrane Centre,MEDLINE,EMBASE,中国生物医学文献数据库(CBM),中文全文期刊数据库(CNKI)和维普中文期刊数据库(VIP)等数据库(截至2007年1月),全面搜集有关复方丹参方制剂(丹参、三七、冰片组成)治疗稳定型心绞痛的随机对照试验。质量评价参照Cochrane handbook4.26提供的标准。[结果]有33篇文献纳入评价。32个研究采用心绞痛症状改善为指标,合并效应量RR(random)及95%可信区间(CI)为1.16(1.10,1.23)。30个研究采用心电图改善为指标,合并效应量RR(random)及95%可信区间为1.38(1.25,1.51)。复方丹参方制剂的不良反应主要表现为轻度的胃部不适,发生率为3.4%,95%CI(2.5%,4.2%)。[结论]根据系统评价结果,提示复方丹参方治疗冠心病心绞痛与硝酸酯类相比,在改善心绞痛症状、改善心肌缺血状态等方面显示出一定的优势,且不良反应少。但由于原始研究质量较低,影响了结论的可靠性,有待高质量研究提供更有力的证据。