北虫草废料酶解液中腺苷和虫草素含量的HPLC分析

测定人工北虫草及废料酶解液中腺苷和虫草素含量,判断酶解过程对腺苷和虫草素的影响。以kromasil 5u C18色谱柱,甲醇-水为流动相,采用梯度洗脱,流速0.8 mL/min,检测波长260 nm,柱温30℃。人工北虫草废料酶解液中腺苷和腺苷的含量为11.24 mg/L和70.40 mg/L。废料酶解液中含有一定量的腺苷和虫草素,具有利用价值。     

虫草活性成分虫草素临床应用研究进展

综述了延缓虫草素快速代谢的三类方法：附加防止脱氨基的侧链基团、与腺苷脱氨抑制剂联合应用、采用纳米材料包埋及其研究进展,指出选用新型材料,采用纳米技术包埋虫草素是解决虫草素快速代谢问题的很好的方法。     

HPLC法同步测定蛹虫草中虫草素及腺苷的含量

目的:采用HPLC法同步测定蛹虫草中虫草素及腺苷的含量。方法:采用SGE protecol C18(5μm,250mm×4.6mm)色谱柱,流动相为乙腈-水(5∶95,V/V),检测波长为260nm,流速1.0m L/min,进样量20μL。结果:虫草素和腺苷分别在16.875~540μg/m L、19.375~620μg/m L范围内呈线性相关,相关系数r分别为0.9998、0.9996。该方法重复性、精密度、稳定性及回收率均符合要求。结论:本法用于同步测定蛹虫草中虫草素及腺苷的含量,具有简便、准确和高效等特点。     

蛹虫草多糖和腺苷药理作用的研究进展

蛹虫草寄生于鳞翅目昆虫蛹及幼虫上,是由子座和菌核组合成的复合体.研究表明,蛹虫草中的虫草多糖具有抗肿瘤、免疫调节、降血糖等药理作用;其中的腺苷具有改善心脑血液循环、防止心律失常和保护肾脏等药理作用.本文将对虫草多糖和腺苷药理作用的研究进展进行综述.     

高效液相色谱法测定虫草发酵菌丝粉中腺苷的含量

本文讨论了高效液相色谱法测定虫草发酵菌丝粉中腺苷含量的方法。实验表明:在ODSHypersil(5μm,125×4mm)色谱柱上,以0.06mol/L磷酸二氢钾-甲醇-四氢呋喃(150:10:1.5)为流动相,检测波长260nm,腺苷的检出限为50.25μg/mL,成功测定了不同批号虫草发酵菌丝粉中腺苷的含量,平均回收率为101%,分析方法灵敏、准确、重现性好。     

蛹虫草多糖提取分离方法的研究进展

蛹虫草含有虫草素、虫草酸、虫草多糖等多种高药理价值的化合物。其中蛹虫草多糖的含量为4%～10%,是其主要有效成分,具有抗癌、抗氧化、增强免疫等作用。本文综述了蛹虫草多糖的提取、分离方法的研究进展,为后续药理研究以及产业化生产提供参考。     

虫草素抗肿瘤作用的机制研究

虫草素(3’-脱氧腺苷),又名虫草菌素,因在人类健康方面具有许多药理和治疗意义而越来越被大众所知。包括抗菌消炎、抗病毒、抗肿瘤、免疫调节等活性,对多种疾病尤其是恶性肿瘤有着明显的治疗作用。虫草素对免疫、肝、肾、心血管系统具有广谱药理潜力。这些年来,研究人员一直对虫草素抗肿瘤的机制不懈地研究,更多的药理活性被发现,抗肿瘤的作用机制也已成为研究热点。本文对虫草素的抗肿瘤作用机制进行综述,为临床应用提供理论基础。     

利用啤酒糟固体发酵产虫草菌素的研究

以啤酒糟为基质,蛹虫草为出发菌株,固态发酵生产虫草菌素。用单因素法和正交实验对培养基中的含水量、碳氮源及其浓度和无机盐进行优化。结果表明:培养基70%含水量、2%淀粉、1%蛋白胨、0.05%KH2PO4和0.05%K2HPO4为最优条件。在优化的发酵条件下,最大虫草素积累量5000.48μg/g,较优化前提高了133.99%。     

虫草素的化学合成进展

虫草素第一个从真菌中分离出来的核苷类抗菌素,它具有多种药理活性作用。在国际市场上,虫草素的价格非常昂贵。因此,许多实验室对虫草素的合成进行了研究。本文对虫草素的化学合成方法进行了全面总结,并作出合适的评价和分析。     

虫草活菌素生物制品

1　项目简介虫草活菌素制品是采用组培技术对冬虫夏草人工培养,经生物技术分离提纯,将其有效成分与蜡状芽孢菌进行配比,再经冷冻干燥制成制剂———冻干制品,该产品是新一代保健品———具有益肾阳、补肺阴、补虚损功效,能够提高机体免疫能力、促进消化、消除酸胀,对抗菌素无效的腹泻有明显的效果,同时有增强记忆、降血脂、镇静催眠、消除腰膝酸痛之功能,对预防动脉硬化肥胖症有一定的作用。2　项目完成程度该项目已完成制品的小样。技术水平,科技含量高,目前正在申请批号。3　生产性投资该项目占地面积需1500～2000ｍ2,包括虫草的培养需100…     

Adenosine and Cordycepin Accelerate Tissue Remodeling Process through Adenosine Receptor Mediated Wnt/β-Catenin Pathway Stimulation by Regulating GSK3b Activity

Adenosine is a cellular metabolite with diverse derivatives that possesses a wide range of physiological roles. We investigated the molecular mechanisms of adenosine and cordycepin for their promoting effects in wound-healing process. The mitochondrial energy metabolism and cell proliferation markers, cAMP responsive element binding protein 1 (CREB1) and Ki67, were enhanced by adenosine and cordycepin in cultured dermal fibroblasts. Adenosine and cordycepin stimulated adenosine receptor signaling via elevated cAMP. The phosphorylation of mitogen-activated protein kinase kinase (MEK) 1/2, mammalian target of rapamycin (mTOR) and glycogen synthase kinase 3 beta (Gsk3b) and Wnt target genes such as bone morphogenetic protein (BMP) 2/4 and lymphoid enhancer binding factor (Lef) 1 were activated. The enhanced gene expression by adenosine and cordycepin was abrogated by adenosine A_2A and A_2B receptor inhibitors, ZM241385 and PSH603, and protein kinase A (PKA) inhibitor H89, indicating the involvement of adenosine receptor A_2A, A_2B and PKA. As a result of Wnt/β-catenin pathway activation, the secretion of growth factors such as insulin-like growth factor (IGF)-1 and transforming growth factor beta (TGFβ) 3 was increased, previously reported to facilitate the wound healing process. In addition, in vitro fibroblast migration was also increased, demonstrating their possible roles in facilitating the wound healing process. In conclusion, our data strongly demonstrate that adenosine and cordycepin stimulate the Wnt/β-catenin signaling through the activation of adenosine receptor, possibly promoting the tissue remodeling process and suggest their therapeutic potential for treating skin wounds.     

Enzyme-assisted extraction of cordycepin and adenosine from cultured Cordyceps militaris and purification by macroporous resin column chromatography

Enzyme-assisted extraction (EAE) coupled with macroporous resin column chromatography (MRCC) was successfully used for the extraction and purification of cordycepin and adenosine from cultured Cordyceps militaris. After optimized by the response surface methodology (RSM), the extraction yields of cordycepin and adenosine were 86.45% and 70.06%, respectively. Subsequently, under the optimal separation conditions on NKA-II resins (loading 1 BV of extraction solution with pH 8.0, and eluting with 4 BVs of 70% ethanol at a flow rate of 3 BV/h), cordycepin (purity of 32.5%), and adenosine (purity of 39.9%) were obtained with an overall recovery rate more than 90%.     

A Systematic Study on the Optimal Nucleotide Analogue Concentration and Rate Limiting Nucleotide of the SARS-CoV-2 RNA-Dependent RNA Polymerase

The current COVID-19 pandemic has highlighted the necessity of more efficient antiviral compounds. The antiviral efficacy of adenosine-based analogs, the main repurposed drugs for SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) inhibition, is mainly assessed through in vitro or cell-free polymerization assays, under arbitrary conditions that do not reflect the physiological environment. We show that SARS-CoV-2 RdRp inhibition efficiency of remdesivir and cordycepin, two common adenosine analogs, is influenced by endogenous adenosine level, and that the current clinically approved concentrations for COVID-19 treatment are suboptimal for effective RdRp inhibition. Furthermore, we identified GTP as the rate-limiting nucleotide of SARS-CoV-2 replication. Our results demonstrate that nucleotide sensitivity of the RdRp complex and competition of nucleoside analog drugs against endogenous concentrations of nucleotides are crucial elements to be considered when designing new SARS-CoV-2 antiviral compounds.     

红树林内生真菌Paecilomyces sp.(Tree1-7)脂肪酸成分分析

采用φ(硫酸)=1%的甲醇溶液酯化法(方法1)和酸碱结合甲酯化法(方法2)分别对红树林内生真菌Paecilomyces sp.(Tree1-7)代谢产物的石油醚部位衍生化,用气相色谱-质谱法分析鉴定了其成分,测定了各成分的相对质量分数。两种方法鉴定出共有主要脂肪酸成分6种,分别为9,12-十八碳二烯酸、棕榈酸、硬脂酸、顺-9-十八碳烯酸、12-甲基-十四烷酸和Cyclopentaneundecanoic acid。其中9,12-十八碳二烯酸为红树林内生真菌Paecilomyces sp.(Tree1-7)石油醚部位的主要成分,其相对质量分数达56%以上。但两种方法所得微量脂肪酸种类存在差异,方法1共检测到13种,其中微量脂肪酸7种,而方法2能检测到17种,其中微量脂肪酸11种。与φ(硫酸)=1%的甲醇溶液酯化法相比,酸碱结合甲酯化法所得脂肪酸成分更为全面。     

紫外可见分光光度法测定虫草素含量

用紫外分光光度测定方法研究了由蛹虫草提取虫草素的含量。先利用紫外可见分光光度计进行标准测定以拟订标准回归公式,再将提取样品利用分光光度计检测。结果得出虫草素的最大吸收波长为259 nm,虫草素吸光度与虫草素浓度间的标准回归方程为Y=65.5143 X＋0.0386（R2=0.9801）,结合回归系数从而获得经验公式：虫草素的浓度C（mol/L）=0.0153×A（其中A为吸光度值）,并且利用该方法测定了几种提取样品的虫草素含量。     

表面活性剂对啶菌噁唑防治番茄灰霉病的增效作用

以番茄灰霉病菌为靶标菌,采用菌丝生长速率法、菌丝干重法和离体叶片法分别测定了有机硅、蓖麻油、吐温、烷基萘磺酸盐、烷基酚聚氧乙烯醚、木质素、氮酮及磷酸酯盐等18个品种的表面活性剂对啶菌(噁)唑的增效作用.结果表明:采用不同的毒力测定方法,表面活性剂与杀菌剂所表现出的协同作用有所差异.其中0201B、NP-7、TX100在生长速率法和菌丝干重法中均表现出对啶菌(噁)唑的增效作用,而离体叶片法中仅有添加Silwet L77的处理可显著提高药剂防效,对菌丝的抑制活性最高达83.69%,而对照的抑制率为52.09%.     

淡紫拟青霉菌剂的研究开发

淡紫拟青霉能够破坏线虫卵里的胚胎和早期幼虫,因此是一种很好的生防因子。该项目从番茄根结线虫卵块中分离出一株淡紫拟青霉PL9410,以农副产品为原料生产出菌剂,并进行了中试与推广。每克成品的淡紫拟青霉活孢子数(CFU)不低于10~8,产品保存期超过18个月,每亩施2kg菌剂,能使虫口密度降低80％～90％,产量增加15％～20％。该菌剂为纯生物制品,对人畜安全,可作为呋喃丹等被禁用的化学农药的替代产品。     

虫草素类似物2’-脱氧腺苷脂质体的制备及表征

虫草素具有多种生物活性,通过与新型药物载体脂质体结合,可解决其临床中的脱氨基作用。2'-脱氧腺苷是虫草素的结构类似物,易制备,具有和虫草素相同的紫外最大吸收波长(259nm),是一种优良的虫草素替代品,研究2'-脱氧腺苷脂质体可为虫草素脂质体的研究提供重要的参考。本文采用逆相蒸发法制得球形、粒径170～350nm、包封率为45%、稳定性良好的2'-脱氧腺苷脂质体。为虫草素脂质体研究提供基础数据。