不同剂量丹参注射液联合泼尼松龙治疗口腔粘膜下纤维性变的临床研究

目的：探讨采用不同剂量的丹参注射液联合波尼松龙治疗口腔粘膜下纤维性病的治疗效果,为今后的治疗提供更多的依据。方法：选择从2010年1月至2013年1月期间在我院口腔科治疗的100例口腔粘膜下纤维性病患者,根据门诊号,随机将患者分为低剂量组、次低剂量组、中剂量组、高剂量组和对照组,每组各20例,低剂量组、次低剂量组、中剂量组、高剂量组,分别使用不同剂量丹参注射液联合波尼松龙治疗,对照组单纯使用波尼松龙治疗,观察治疗一个疗程后患者口腔粘膜情况及张口度。结果：中剂量组和高剂量组情况改善要明显好于低剂量、次低剂量组、对照组,差异具有显著性（P〈0．05）。结论：丹参注射液联合泼尼松龙治疗口腔粘膜下纤维性病疗效令人满意,其中低剂量丹参注射液便有效果,一定范围内剂量越高,疗效越好,值得在临床推f,     

丹参雄性不育系Sh-B的鉴定与花粉发育过程的解剖学研究

在显微水平上对新发现的丹参雄性不育系Sh-B花药发育过程进行了解剖学观察,并对其花粉活力和结实率进行了鉴定。结果显示:根据花器官及花药的形态、大小以及花丝的长度,可以将Sh-B不育株分为3个不育类型,即Sh-B1、Sh-B2和Sh-B3。这3种不育类型均属于雄性不育,其花丝不到正常可育株的1/2,花药干瘪而瘦小,内无花粉粒或花粉无活力;其根、茎、叶以及种子形态结构与正常可育植株基本相似。产生雄性不育的主要原因有:花粉囊药室内壁纤维层加厚,影响花药壁开裂;小孢子母细胞周围不产生胼胝质或产生的胼胝质很少;绒毡层细胞延迟解体;花粉粒畸形。在其花药发育的小孢子母细胞时期、四分体形成前期、单核期、双核期均可能产生雄性不育的小孢子或花粉粒。     

Determination of phenolic compounds and diterpenes in roots of Salvia miltiorrhiza and Salvia przewalskii by two LC–MS tools: Multi-stage and high resolution tandem mass spectrometry with assessment of antioxidant capacity

Comparative phytochemical analyses of hydroalcoholic (50% EtOH) extracts from roots of S. miltiorrhiza (SM) and S. przewalskii (SP) were performed using two complementary LC–MS systems: the first system HPLC-DAD-MSⁿ an ion trap mass spectrometer and the second system consisted high resolution MS/MS Orbitrap mass spectrometer. The individual compounds were identified using a previously published approach via comparison of the exact molecular masses, mass spectra and retention times to those of standard compounds, online available databases and literature data. Moreover, the determination of antioxidative activities of extracts by DPPH and FRAP methods was carried out. Analysis allowed to identify 39 chemical compounds in extracts from both species. Extract from root of SP differs from SM in the presence of several metabolites such as: przewalskinic acid and their derivatives, przewaquinone C, przewaquinonate A, glycosides of rosmarinic acid, methyltanshinonate, whereas tanshinones, salvianolic acids and lithospermic acids occurred in both species. Moreover, it was shown that hydroalcoholic extract from roots of SM exerted stronger antioxidant properties in a FRAP test (max. 323.92 μM Fe²⁺/L) and in DPPH test (max. 78.64 nM TE) in comparison with SP extract.     

连作对丹参生长的障碍效应

通过对1～4年不同连作年限地块丹参生长发育情况进行的连续调查分析,结果表明,连作严重危害丹参生长,主要表现在6～9月,丹参生长期的枯苗率大幅度上升,地上、地下部生长量下降,根系数量、直径和长度减小,产世降低。同时由于有效成分含量降低,根系外观畸形,造成商品率下降,从而影响丹参质量。通过对土壤pH测定发现．丹参连作障碍可能与土壤酸性增加有关。     

ESI-Q-TOF-MS直接进样法比较陕西五个不同地区丹参的化学成分差异

该研究采用ESI-Q-TOF-MS直接进样分析法对陕西商州、洛南、大荔、丹凤和铜川等五个地区丹参的化学成分进行了比较分析,通过丹参水溶性和脂溶性成分的质谱丰度差异评价了不同地区丹参化学成分含量的变化,对不同产地丹参的化学成分进行了鉴定,综合分析选出了最优的丹参种植产地。结果表明:五地丹参均含有丹参酮ⅡA、隐丹参酮、丹参酮Ⅰ、二氢丹参酮、丹参酚酮、次丹参醌、丹参醌Ⅱ等9种脂溶性化学成分和丹参素、咖啡酸、阿魏酸、迷迭香酸、原紫草酸、紫草酸、原儿茶醛、丹酚酸A、丹酚酸B等9种水溶性化学成分,其中丹参素钠、紫草酸、丹酚酸B和隐丹参酮的含量普遍较高,质谱丰度均大于30%。但是,随着种植环境不同,不同地区丹参的化学成分含量差异很大,如商州产丹参中含有较多具有生物活性的丹参酮类物质,其中丹参酮Ⅰ含量远远高于其他四个产地,质谱丰度达到72.6%,而其他地区丹参酮Ⅰ的质谱丰度为1.8%～11.3%。这表明丹参质量按照地区排序为商州>铜川>大荔>洛南>丹凤。该方法为中药药材质量评价提供了科学、可靠、便捷的途径,为药材规格等级的制定提供了新途径,为陕西丹参种植区域的选择提供了重要信息。     

丹参悬浮培养细胞原生质体的制备和活力检测

对丹参悬浮培养细胞原生质体制备条件进行了研究,并利用FDA染色和钙离子荧光探针Fluo-3/AM装载对制备得到的原生质体的活力和功能进行了检测。丹参悬浮培养细胞原生质体的制备条件为:悬浮培养细胞酶解的适宜酶液组合为纤维素酶1.5%、果胶酶0.3%和离析酶0.5%;适宜的甘露醇浓度为0.4 mol/L;酶解时间为12 h;在600 r/min转速下离心5 min收集,纯化得到原生质体,其产量为1.1×106/g FW,FDA检测显示其活力为95%以上,荧光探针Fluo-3/AM可成功装载到原生质体中。     

内蒙古欧李果肉和果仁中营养成分分析

对内蒙古欧李果肉及果仁中营养成分进行研究,结果表明,欧李中维生素、矿物质、蛋白质等营养成分丰富,其中VC的质量分数为590.1 mg.kg-1,远远高于苹果、草莓、葡萄、柑橘、西红柿等水果的VC含量;矿物质元素Ca质量分数在果肉中达428.1 mg.kg-1,是其它水果Ca含量的2~10倍;果仁中达3.743 g.kg-1,其含量与杏仁中相当;蛋白质含量丰富,氨基酸种类齐全,必需氨基酸在果肉和果仁中所占比例达氨基酸总量的1/3以上,为以后的研究开发利用提供了理论依据。     

Matrix metalloproteinase-9 induces cardiac fibroblast migration, collagen and cytokine secretion: inhibition by salvianolic acid B from Salvia miltiorrhiza

Cardiac fibroblasts play the key role in cardiac function and matrix metalloproteinases-9 (MMP-9) is a well known contributor to the development of myocardial remodeling. However, the direct regulation of MMP-9 on the function of cardiac fibroblasts and the underlying mechanism are far from elucidation. In the present research, recombinant protein encoding catalytic domain of MMP-9 (MMP-9 CD) was constructed and the function of neonatal cardiac fibroblasts was investigated by cell proliferation assay, migration assay, picrosirius red assay, multiplex cytokine assay and fibroblast phenotype detection. 200 nM MMP-9 CD stimulated cardiac fibroblasts migration (169.4±22.5% versus 100±0%, p<0.01), increased collagen synthesis (1.5±0.2 fold, p<0.05), up-regulated the secretion of ICAM (574.0±40.1 versus 268.5±8.6pg/ml, p<0.01), TNF-α (192.6±11.0 versus 14.4±1.8pg/ml, p<0.001), IL-6 (1500.9±70.2 versus 323.4±40.6pg/ml, p<0.001) and sVCAM-1 (30.3±4.3 versus 7.0±0.1 pg/ml, p<0.05) and down-regulated VEGF (436.5±148.9 versus 1034.3±28.1 pg/ml, p<0.05) significantly with modest effects on proliferation. Accompanying with these regulations, transition of fibroblasts to myofibroblast was confirmed by immunofluorescent stain of α-smooth muscle actin (α-SMA) with MMP-9 CD treatment. Furthermore, salvianolic acid B (SalB) inhibited the effects of MMP-9 CD significantly. In conclusion, our results provide evidence for a direct influence of MMP-9 on cardiac fibroblast migration, collagen and cytokine secretion, which can be attenuated by SalB.     

放线菌Act12与腐植酸钾配施对丹参生长及其根域微生态的影响

探讨生防放线菌菌剂与腐植酸钾配施对丹参生长及其根域微生态的影响。以常规移栽处理为对照,研究小区试验中放线菌菌剂与腐植酸钾不同配施比例下对丹参生长、产量及抗根结线虫侵染的影响;并采用稀释平皿涂抹法测定丹参根区土壤、根表土壤、根外土壤及根系中细菌(B)、真菌(F)与放线菌(A)的数量,同时对优势细菌、真菌和放线菌进行了分子生物学鉴定,研究放线菌菌剂与腐植酸钾配施处理下丹参根域微生态变化。研究结果表明:1配施能增强菌剂对丹参的促生效果。菌剂与腐植酸钾配施T20处理丹参出苗率较对照提高8.7%,收获时的死亡率较对照减少39.0%;茎叶鲜质量、根鲜质量、单株根鲜质量、根干质量以及单株根干质量分别较对照增加6.1%、28.6%、11.1%、36.3%以及9.0%。2可以调整丹参植株根域土壤微生态平衡,降低有害微生物数量,增加有益微生物数量,改善微生物区系。在丹参根表土壤中,菌剂与腐植酸钾配施处理B/A值较对照降低78.4%,A/F值较对照增加95.0%。在丹参根系内,菌剂与腐植酸钾配施处理细菌数量较对照增加195.0%,未检测到真菌和放线菌存在。3在放线菌处理丹参根区、根表土壤中,有6株优势菌可能对丹参生长及抗病有益:3株优势细菌分别为硝基愈疮木胶节杆菌(Arthrobacter nitroguajacolicus)、放射型根瘤菌(Rhizobium radiobacter)和弗雷德里克斯堡假单胞菌(Pseudomonas frederiksbergensis);3株优势放线菌分别为淀粉酶产色链霉菌(Streptomyces diastatochromogenes)、砖红链霉菌(S.lateritius)和卡伍尔链霉菌(S.cavourensis)。有2株优势菌疑为有害微生物:优势细菌为耐寒短杆菌(Brevibacterium frigoritolerans),优势放线菌为肿痂链霉菌(S.turgidiscabies)。这2种菌对其他作物的有害作用已有报道。4对丹参根结线虫侵染有强烈抑制作用,可使田间根结线虫侵染率降低49.3%。生防放线菌与腐植酸钾配施处理后能明显促进丹参生长,提高丹参产量及抗病虫能力,调节丹参根域微生态平衡。     

Molecular Identification and Diversity of ‘Candidatus Phytoplasma solani’ Associated with Red‐leaf Disease of Salvia miltiorrhiza in China

Reddening disease has recently been threatening Salvia miltiorrhiza in China, ranging from 30 to 50%. The main symptoms observed, such as plant stunting, inflorescence malformation, leaf reddening, fibrous roots browning, skin blackening and eventually root rot, are typically associated with phytoplasma infection. The presence of phytoplasmas was demonstrated through phytoplasma‐specific PCR, with the expected amplification (1.8 kb) from symptomatic S. miltiorrhiza plants from Shangluo, Shangzhou and Luonan fields in Shaanxi Province of China. The sequences of 16S rRNA, tuf, secY and vmp1 genes amplified from LN‐1 phytoplasma shared the closest homologies of 99%, 100%, 99% and 98% with those of the reference strain Candidatus Phytoplasma solani (subgroup 16SrXII‐A), respectively. The phylogenetic trees showed that LN‐1 phytoplasma clustered with the members of 16SrXII‐A group, including Ca. P. solani. Computer‐simulated restriction fragment length polymorphism analysis further supported this classification. Diversity analysis showed that all ‘Ca. P. solani’ strains identified from the three different regions examined shared 100% identical 16S rRNA, tuf, secY and vmp1 nucleotide sequences. To the best of our knowledge, this is the first report of phytoplasma infecting the medicinal plant of S. miltiorrhiza. The results demonstrate that ‘Ca. P. solani’ is the presumptive aetiological agent of S. miltiorrhiza reddening disease in China.